Cephem compounds

ABSTRACT

The invention relates particularly to syn compounds of the formula: &lt;IMAGE&gt;   wherein R2 is hydrogen or an aliphatic hydrocarbon residue which may be substituted with halogen, carboxy or esterified carboxy, R5 is carboxy or functionally modified carboxy and R6 is amino or protected amino, and pharmaceutically acceptable salt thereof, their preparation and their pharmaceutical uses.

This is a division of application Ser. No. 886,340, filed Mar. 14, 1978and of application Ser. No. 302,668 filed Sept. 15, 1981 now U.S. Pat.No. 4,427,674.

This invention relates to new cephem and cepham compounds. Moreparticularly, it relates to new 7-substituted-3-cephem(orcepham)-4-carboxylic acid, its pharmaceutically acceptable salt andpharmaceutically acceptable bioprecursor thereof, which haveantimicrobial activities, and processes for preparation thereof, tointermediate for preparing the same and processes for preparationthereof, and to pharmaceutical composition comprising the same andmethods of using the same prophylactically and therapeutically fortreatment of infectious diseases in human being and animals.

Accordingly, the objects of this invention are to provide:

new 7-substituted-3-cephem(or cepham)-4-carboxylic acid, itspharmaceutically acceptable salt and pharmaceutically acceptablebioprecursor thereof, which exhibit excellent antimicrobial activitiesagainst a wide variety of pathogenic microorganisms including Gramnegative and Gram positive bacteria,

processes for preparation of the same,

pharmaceutical composition comprising one of the same as an activeingredient, and

a method of using the same prophylactically and therapeutically fortreatment of infectious diseases caused by pathogenic microorganisms inhuman being and animals; and further

intermediate to be used for preparation of pharmaceutically active7-substituted-3-cephem(or cepham)-4-carboxylic acid, itspharmaceutically acceptable salt or pharmaceutically acceptablebioprecursor thereof, and

methods for preparation of the same.

The cephem and cepham compounds provided by this invention can berepresented by the formula (I): ##STR2## wherein R¹ is thiadiazolyl,thiazolyl of the formula: ##STR3## in which R⁶ is amino or protectedamino, or haloacetyl, A is methylene or a group of the formula: ##STR4##in which R² is hydrogen or an aliphatic hydrocarbon residue which may besubstituted with halogen, carboxy or esterified carboxy,

R³ is hydrogen or lower alkyl,

R⁴ is hydrogen, halogen, lower alkyl or a group of the formula: --O--R⁷in which R⁷ is hydrogen, lower alkyl or acyl,

R⁵ is carboxy or functionally modified carboxy, and

the dotted line represents 3-cephem and cepham nuclei, inclusively,provided that

(i) R⁴ is hydrogen, halogen or a group of the formula: --O--R⁷ in whichR⁷ is as defined above, when R³ is hydrogen,

(ii) R⁴ is lower alkyl, when R³ is lower alkyl,

(iii) A is a group of the formula: ##STR5## in which R² is as definedabove, when R¹ is thiadiazolyl or thiazolyl of the formula: ##STR6## inwhich R⁶ is as defined above, and (iv) the dotted line represents3-cephem nucleus and R⁴ is hydrogen, halogen, lower alkyl or --OR⁷ inwhich R⁷ is lower alkyl, when R¹ is haloacetyl.

It is to be noted that the cephem and cepham compounds (I) asillustrated above include a compound useful as an antimicrobial agentand also a compound useful as an intermediate for preparing the aboveantimicrobial agent, particularly as illustrated below.

The compound useful as an antimicrobial agent can be represented by theformula (I'): ##STR7## wherein R_(a) ¹ is thiadiazolyl or thiazolyl ofthe formula: ##STR8## in which R⁶ is as defined above, and R², R³, R⁴and R⁵ are each as defined above.

On the other hand, the compound useful as an intermediate for preparingthe above compound (I') can be represented by the formula (I"): ##STR9##wherein R_(b) ¹ is haloacetyl,

R_(a) ⁴ is hydrogen, halogen, lower alkyl or a group of the formula:--O--R⁷ in which R⁷ is lower alkyl, and R³, R⁵ and A are each as definedabove.

And further, it is to be noted that the compound (I') where R_(a) ¹ isthiazolyl of the formula: ##STR10## in which R⁶ is protected amino, R⁴is a group of the formula: --O--R⁷ in which R⁷ is hydrogen or acyland/or R⁵ is functionally modified carboxy is also useful as anintermediate for preparing the more active compound as explained below.

Accordingly, the more preferred active compound can be represented bythe formula (I'"): ##STR11## wherein R_(c) ¹ is thiadiazolyl orthiazolyl of the formula: ##STR12## and R², R³ and R_(a) ⁴ are each asdefined above.

The terms and definitions described in this specification and claims areillustrated as follows.

(a) Partial structure of the formula: ##STR13## is intended to mean bothof the geometric formulae: ##STR14##

The geometry of the formula (S) is referred to as "syn" and anotherformula (A) is referred to as "anti".

Accordingly, one isomer of the compound having the partial structureshown by the above formula (S) is referred to as "syn isomer" andanother isomer of the compound having the alternative one shown by theabove formula (A) is referred to as "anti isomer", respectively.

From the view point of structure-activity relationship, it is to benoted that a syn isomer of the compound (I') tends to be of much higherantimicrobial activity than the corresponding anti isomer, andaccordingly the syn isomer of the compound (I') is more preferableantimicrobial agent than the corresponding anti isomer in theprophylactic and therapeutic value.

(b) The thiazolyl group of the formula: ##STR15## (wherein R⁶ is asdefined above) is well known to lie in tautomeric relation with athiazolinyl group of the formula: ##STR16## (wherein R^(6') is imino orprotected imino).

The tautomerism between the said thiazolyl and thiazolinyl groups can beillustrated by the following equilibrium: ##STR17## (wherein R⁶ andR^(6') are each as defined above).

Accordingly, it is to be understood that both of the said groups aresubstantially the same, and the tautomers consisting of such groups areregarded as the same compounds, especially in the manufacturingchemistry. Therefore, both of the tautomeric forms of the compoundshaving such groups in their molecule are included in the scope of thisinvention and designated inclusively with one expression "thiazolyl" andrepresented by the formula: ##STR18## (wherein R⁶ is as defined above)only for the convenient sake throughout this specification and claims.

(c) It is well known that the 3-hydroxy-3-cephem compound having thepartial structure of the formula: ##STR19## lies in a tautomericrelation with the 3-oxo-cepham compound of the formula: ##STR20## eachof which is referred to as the enol- or keto-tautomer, and that theenol-tautomer is usually the stabilized one.

Accordingly, both of the compounds having such tautomeric structures areincluded within the same scope of the compound, and therefore, thestructure and nomenclature of such tautomers are expressed inclusivelywith one expression of the stabilized enol tautomer, i.e.3-hydroxy-3-cephem compound throughout this specification and claims.

In the above and subsequent descriptions of this specification, suitableexamples and illustration of the various definitions which thisinvention intends to include within the scope thereof are explained indetail as follows.

The term "lower" is used to intend a group having 1 to 6 carbon atoms,unless otherwise provided.

"Thiadiazolyl" for R¹ may be 1,2,3-thiadiazolyl (e.g.1,2,3-thiadiazol-4-yl or 1,2,3-thiadiazol-5-yl), 1,3,4-thiadiazolyl or1,2,4-thiadiazolyl, preferably 1,2,3-thiadiazolyl, and more preferably1,2,3-thiadiazol-4-yl.

"Aliphatic hydrocarbon residue" for R² may include a monovalent radicalof a saturated or unsaturated, and straight, branched or cyclicaliphatic hydrocarbon, and particularly may include alkyl, alkenyl,alkynyl, cycloalkyl and the like, the details of which are explainedbelow.

"Alkyl" may include a residue of straight or branched alkane having 1 to12 carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, t-butyl, pentyl, neopentyl, hexyl, heptyl, octyl, nonyl,decyl, undecyl, dodecyl and the like, preferably lower alkyl, and morepreferably the one having 1 to 4 carbon atoms.

"Alkenyl" may include a residue of a straight or branched alkene havingup to 12 carbon atoms, preferably lower alkenyl such as vinyl, allyl,1-propenyl, isopropenyl, butenyl, isobutenyl, pentenyl, hexenyl and thelike, and more preferably the ones having up to 4 carbon atoms.

"Alkynyl" may include a residue of a straight or branched alkyne havingup to 12 carbon atoms, preferably lower alkynyl such as ethynyl,propargyl, 1-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl,2-pentynyl, 1-pentynyl, 5-hexynyl and the like, and more preferably theones having up to 4 carbon atoms.

"Cycloalkyl" may include a residue of a cycloalkane having up to 8carbon atoms, preferably lower cycloalkyl such as cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl, and more preferably cyclohexyl.

These aliphatic hydrocarbon residues may be substituted with halogenatom(s), carboxy or esterified carboxy group(s). Accordingly, "aliphatichydrocarbon residue substituted with halogen atom(s), carboxy oresterified carboxy group(s)" can also be alternatively expressed as"halogen-substituted aliphatic hydrocarbon residue","carboxy-substituted aliphatic hydrocarbon residue" and "esterifiedcarboxy-substituted aliphatic hydrocarbon residue", respectively, whichmay include more particularly halo-alkyl, alkenyl, alkynyl andcycloalkyl; carboxy-alkyl, alkenyl, alkynyl and cycloalkyl; andesterified carboxyalkyl, alkenyl, alkynyl and cycloalkyl, respectively.

Suitable examples of the "halogen" may include chlorine, bromine, iodineand fluorine; suitable examples of the "esterified carboxy" may bealkoxycarbonyl or the like; and preferred examples of the "alkyl","alkenyl", "alkynyl", "cycloalkyl" and alkyl moiety of the"alkoxycarbonyl" are the corresponding "lower" ones as mentioned above.

Preferred examples of the "halo-alkyl, alkenyl, alkynyl and cycloalkyl"may be chloromethyl, bromomethyl, iodomethyl, fluoromethyl,trichloromethyl, trifluoromethyl, 2-chloroethyl, 1,2-dichloroethyl,2,2,2-trifluoroethyl, 3-chloropropyl, 4-iodobutyl, 5-fluoropentyl,6-bromohexyl, 3-fluoroallyl, 3-chloropropargyl, 4-fluorocyclohexyl, orthe like.

Preferred examples of the "carboxy-alkyl, alkenyl, alkynyl andcycloalkyl" may be carboxymethyl, 1-carboxyethyl, 2-carboxyethyl,1-carboxypropyl, 3-carboxypropyl, 4-carboxybutyl, 5-carboxypentyl,6-carboxyhexyl, 1-carboxyisopropyl, 1-ethyl-1-carboxyethyl,2-methyl-2-carboxypropyl, 3-carboxyallyl, 3-carboxypropargyl,4-carboxycyclohexyl, or the like.

Preferred examples of the "esterified carboxyalkyl, alkenyl, alkynyl andcycloalkyl" may be lower alkoxycarbonyl(lower)alkyl (e.g.methoxycarbonylmethyl, ethoxycarbonylmethyl, propoxycarbonylmethyl,t-butoxycarbonylmethyl, 2-ethoxycarbonylethyl, 2-ethoxycarbonylpropyl,4-ethoxycarbonylbutyl, 1-t-butoxycarbonylisopropyl,1-t-butoxycarbonyl-1-methylpropyl, 4-t-butoxycarbonylbutyl,5-t-butoxycarbonylpentyl, 6-butoxycarbonylhexyl, etc.), loweralkoxycarbonyl(lower)alkenyl (e.g. 3-methoxycarbonylallyl, etc.), loweralkoxycarbonyl(lower)alkynyl (e.g. 3-methoxycarbonylpropargyl, etc.),lower alkoxycarbonyl(lower)cycloalkyl (e.g. 4-methoxycarbonylcyclohexyl,etc.) or the like, and more preferably lower alkoxycarbonylmethyl asexemplified above.

"Lower alkyl" for R³, R⁴ and R⁷ is to be referred to those asexemplified in the term of the aliphatic hydrocarbon residue for R²,preferably may be the ones having up to 4 carbon atoms and morepreferably methyl.

"Halogen" for R⁴ may be chlorine, bromine, iodine or fluorine, andpreferred one is chlorine or bromine.

"Acyl" for R⁷ may be lower alkanoyl (e.g. formyl, acetyl, propionyl,butyryl, isobutyryl, isovaleryl, pivaroyl, etc.), aroyl (e.g. benzoyl,etc.), lower alkanesulfonyl (e.g. mesyl, ethanesulfonyl,1-methylethanesulfonyl, propanesulfonyl, butanesulfonyl, etc.),arenesulfonyl (e.g. benzenesulfonyl, tosyl, etc.) or the like.

"Protective group" in the "protected amino" for R⁶ may be theconventional N-protective group such as substituted or unsubstitutedar(lower)alkyl (e.g. benzyl, benzhydryl, trityl, 4-methoxybenzyl,3,4-dimethoxybenzyl, etc.), halo(lower)alkyl (e.g. trichloromethyl,trichloroethyl, trifluoromethyl, etc.), tetrahydropyranyl, substitutedphenylthio, substituted alkylidene, substituted aralkylidene,substituted cycloalkylidene, acyl, or the like.

Suitable acyl for the protective group may be substituted orunsubstituted lower alkanoyl (e.g. formyl, acetyl, chloroacetyl,trifluoroacetyl, etc.), substituted or unsubstituted loweralkoxycarbonyl (e.g. methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl,1-cyclopropylethoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl,t-butoxycarbonyl, pentyloxycarbonyl, t-pentyloxycarbonyl,hexyloxycarbonyl, trichloroethoxycarbonyl, 2-pyridylmethoxycarbonyl,etc.), substituted or unsubstituted ar(lower)alkoxycarbonyl (e.g.benzyloxycarbonyl, benzhydryloxycarbonyl, 4-nitrobenzyloxycarbonyl,etc.), lower cycloalkoxycarbonyl (e.g. cyclopentyloxycarbonyl,cyclohexyloxycarbonyl, etc.), 8-quinolyloxycarbonyl, succinyl,phthaloyl, or the like.

And further, the reaction product of a silan, boron, aluminium orphosphorus compound with the amino group may also be included in theprotective group. Suitable examples of such compounds may betrimethylsilyl chloride, trimethoxysilyl chloride, boron trichloride,butoxyboron dichloride, aluminum trichloride, diethoxy aluminumchloride, phosphorus dibromide, phenylphosphorus dibromide, or the like.

"Functionally modified carboxy" for R⁵ may be an ester, amide or thelike.

Suitable examples of the ester may be

alkyl ester (e.g. methyl ester, ethyl ester, propyl ester, isopropylester, butyl ester, isobutyl ester, t-butyl ester, pentyl ester,t-pentyl ester, hexyl ester, heptyl ester, octyl ester,1-cyclopropylethyl ester, etc.);

alkenyl ester (e.g. vinyl ester, allyl ester, etc.);

alkynyl ester (e.g. ethynyl ester, propynyl ester, etc.);

alkoxyalkyl ester (e.g. methoxymethyl ester, ethoxymethyl ester,isopropoxymethyl ester, 1-methoxyethyl ester, 1-ethoxyethyl ester,etc.);

alkylthioalkyl ester (e.g. methylthiomethyl ester, ethylthiomethylester, ethylthioethyl ester, isopropylthiomethyl ester, etc.);

haloalkyl ester (e.g. 2-iodoethyl ester, 2,2,2-trichloroethyl ester,etc.);

alkanoyloxyalkyl ester (e.g. acetoxymethyl ester, propionyloxymethylester, butyryloxymethyl ester, valeryloxymethyl ester, pivaloyloxymethylester, hexanoyloxymethyl ester, 2-acetoxyethyl ester,2-propionyloxyethyl ester, palmitoyloxymethyl ester, etc.);

alkanesulfonylalkyl ester (e.g. mesylmethyl ester, 2-mesylethyl ester,etc.);

substituted or unsubstituted aralkyl ester (e.g. benzyl ester,4-methoxybenzyl ester, 4-nitrobenzyl ester, phenethyl ester, tritylester, benzhydryl ester, bis(methoxyphenyl)methyl ester,3,4-dimethoxybenzyl ester, 4-hydroxy-3,5-di-t-butylbenzyl ester, etc.);

substituted or unsubstituted aryl ester (e.g. phenyl ester, tolyl ester,t-butylphenyl ester, xylyl ester, mesityl ester, cumenyl ester, salicylester, etc.);

an ester with a silyl compound such as trialkylsilyl compound,dialkylalkoxysilyl compound or trialkoxysilyl compound, for example,trialkylsilyl ester (e.g. trimethyl silyl ester, triethylsilyl ester,etc.), dialkylalkoxy silyl ester (e.g. dimethylmethoxysilyl ester,dimethylethoxysilyl ester, diethylmethoxysilyl ester, etc.) ortrialkoxysilyl ester (e.g. trimethoxysilyl ester, triethoxysilyl ester,etc.) or the like.

With regard to the terms "protected amino" for R⁶ and "functionallymodified carboxy" for R⁵, it is to be understood that these groups bearthe meaning not only in synthetic manufacture of the object compound bychemical process(es), but also in physiological and pharmaceuticalproperties of the object compound per se.

That is, in the meaning of the synthetic manufacture, free amino groupfor R⁶ and/or free carboxy group for R⁵ may be transformed into the"protected amino" and/or "functionally modified carboxy" as mentionedabove before conducting the process(es) for preventing any possibleundesired side reaction(s), and the "protected amino" and/or"functionally modified carboxy" group in the resultant compound may betransformed into free amino and/or carboxy group after the reaction isconducted. This will be apparent from the explanation of the processesin the following.

On the other hand, in the meaning of the physiological andpharmaceutical properties of the object compound, the compound bearingthe "protected amio" and/or "functionally modified carboxy" group isoptionally used for improving the properties such as solubility,stability, absorbability, toxicity of the particularly active objectcompound bearing the free amino and/or carboxy group.

Suitable "pharmaceutically acceptable salt" of the object compound (I')may be conventional non-toxic salt, and may include a salt with aninorganic base or acid, for example, a metal salt such as an alkalimetal salt (e.g. sodium salt, potassium salt, etc.) and an alkalineearth metal salt (e.g. calcium salt, magnesium salt, etc.), ammoniumsalt, an inorganic acid salt (e.g. hydrochloride, hydrobromide, sulfate,phosphate, carbonate, bicarbonate, etc.), a salt with an organic base oracid, for example, an amine salt (e.g. trimethylamine salt,triethylamine salt, pyridine salt, procaine salt, picoline salt,dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt,N-methylglucamine salt, diethanolamine salt, triethanolamine salt,tris(hydroxymethylamino)methane salt, phenethylbenzylamine salt, etc.),an organic caboxylic or sulfonic acid salt (e.g. acetate, maleate,lactate, tartrate, mesylate, benzenesulfonate, tosylate, etc.), a basicor acidic amino acid salt (e.g. arginine salt, aspartic acid salt,glutamic acid salt, lysine salt, serine, etc.) and the like.

It is well known in the pharmaceutical field that the active drug, whenit has any undesired physiological or pharmaceutical property such assolubility, stability, absorbability, etc., is converted into modifiedderivative thereof for improving such undesired properties, and thensaid derivative, upon administration to a patient, exhibits the activeefficacy by being converted into the body to the parent drug. In thismeaning, the term "pharmaceutically acceptable bioprecursor" usedthroughout this specification and claim is intended to fundamentallymean all of the modified derivatives, which have structural formulaedifferent from those of the active compounds of this invention, but areconverted in the body to the active compounds of this invention uponadministration, and also to mean the derivatives which are sometimesderived physiologically from the compounds of this invention in the bodyand exhibit antimicrobial efficacy.

The compounds (I) of this invention can be prepared by processes asshown in the following scheme. ##STR21## wherein R_(a) ² is an aliphatichydrocarbon residue which may be substituted with halogen, carboxy oresterified carboxy,

R_(b) ⁴ is halogen,

R_(a) ⁵ is esterified carboxy,

R_(b) ⁵ is functionally modified carboxy,

R_(a) ⁶ is protected amino,

R_(a) ⁷ is acyl, and

R¹, R_(a) ¹, R_(b) ¹, R², R³, R⁴, R_(a) ⁴, R⁵ and A are each as definedabove.

The above processes will be explained in detail in the following.

Process A: N-Acylation

A compound (I) and its salt can be prepared by reacting a7-amino-3-cephem (or cepham) compound (II), its reactive derivative atthe amino or a salt thereof with a carboxylic acid (III), its reactivederivative at the carboxy or a salt thereof according to a conventionalmanner of so-called amidation reaction well known in β-lactam chemistry.

The starting compound (III) includes both of known and new ones, and thenew compound (III) can be prepared according to the methods as explainedhereinafter in this specification.

Suitable reactive derivative at the amino group of the compound (II) mayinclude a conventional reactive derivative as used in a wide variety ofamidation reaction, for example, isocyanato, isothiocyanato, aderivative formed by the reaction of a compound (II) with a silylcompound (e.g. trimethylsilylacetamide, bis(trimethylsilyl)acetamide,etc.), with an aldehyde compound (e.g. acetaldehyde, isopentaldehyde,benzaldehyde, salicylaldehyde, phenylacetaldehyde, p-nitrobenzaldehyde,m-chlorobenzaldehyde, p-chlorobenzaldehyde, hydroxynaphthoaldehyde,furfural, thiophenecarboaldehyde, etc., or the corresponding hydrate,acetal, hemiacetal or enolate thereof), with a ketone compound (e.g.acetone, methyl ethyl ketone, methyl isobutyl ketone, acetylacetone,ethyl acetoacetate, etc., or the corresponding ketal, hemiketal orenolate thereof), with phosphorus compound (e.g. phosphorus oxychloride,phosphorous chloride, etc.), or with a sulfur compound (e.g. thionylchloride, etc.), and the like.

Suitable salt of the compound (II) may be referred to the one asexemplified for the compound (I).

Suitable reactive derivative at the carboxy group of the compound (III)may include, for example, an acid halide, an acid anhydride, anactivated amide, an activated ester, and the like, and preferably acidhalide such as acid chloride, acid bromide; a mixed acid anhydride withan acid such as substituted phosphoric acid (e.g. dialkylphosphoricacid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoricacid, halogenated phosphoric acid, etc.), dialkylphosphorous acid,sulfurous acid, thiosulfuric acid, sulfuric acid, alkylcarbonic acid,aliphatic carboxylic acid (e.g. pivalic acid, pentanoic acid,isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.),aromatic carboxylic acid (e.g. benzoic acid, etc.); a symmetrical acidanhydride; an activated acid amide with imidazole, 4-substitutedimidazole, dimethylpyrazole, triazole or tetrazole; an activated ester(e.g. cyanomethyl ester, methoxymethyl ester, dimethylaminomethyl ester,vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenylester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenylester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester,p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridylester, piperidyl ester, 8-quinolyl thioester, an ester with a N-hydroxycompound such as N,N-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone,N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole,1-hydroxy-6-chlorobenzotriazole, etc.), and the like.

The suitable reactive derivatives of the compounds (II) and (III) can beoptionally selected from the above according to the kind of thecompounds (II) and (III) to be used practically, and to the reactionconditions.

Suitable salt of the compound (III) may include a salt with an inorganicbase such as alkali metal salt (e.g. sodium salt, potassium salt, etc.)and an alkaline earth metal salt (e.g. calcium salt, magnesium salt,etc.), a salt with an organic base such as tertiary amine (e.g.trimethylamine salt, triethylamine salt, N,N-dimethylaniline salt,pyridine salt, etc.), a salt with an inorganic acid (e.g. hydrochloride,hydrobromide, etc.) and the like.

The reaction is usually carried out in a conventional solvent such aswater, acetone, dioxane, acetonitrile, chloroform, benzene, methylenechloride, ethylene chloride, tetrahydrofuran, ethyl acetate,N,N-dimethylformamide, pyridine or any other solvent which does notadversely influence to the reaction, or an optional mixture thereof.

When the acylating agent (III) is used in a form of free acid or salt inthis reaction, the reaction is preferably carried out in the presence ofa condensing agent such as a carbodiimide compound (e.g.N,N'-dicyclohexylcarbodiimide,N-cyclohexyl-N'-morpholinoethylcarbodiimide,N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide,N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide,N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide, etc.), a bisimidazolidecompound (e.g. N,N'-carbonylbis(2-methylimidazole), etc.), an iminecompound (e.g. pentamethyleneketene-N-cyclohexylimine,diphenylketene-N-cyclohexylimine, etc.), an olefinic or acetylenic ethercompound (e.g. ethoxyacetylene, β-chlorovinylethyl ether, etc.),1-(4-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole,N-ethylbenzisoxazolium salt, N-ethyl-5-phenylisoxazolium-3'-sulfonate, aphosphorus compound (e.g. polyphosphoric acid, trialkyl phosphite, ethylpolyphosphate, isopropyl polyphosphate, phosphorus oxychloride,phosphorus trichloride, diethylchlorophosphite, orthophenylenechlorophosphite, etc.), thionyl chloride, oxalyl chloride, Vilsmeierreagent prepared by the reaction of dimethylformamide with thionylchloride, phosphorus oxychloride, phosgene or the like.

With regard to the geometry of the compound (I) wherein A is a group ofthe formula: ##STR22## [hereinafter referred to as "oxyimino compound"(I)] produced by this process, it is to be noted that there seems to bestereoselectivity between syn and anti isomers, as explained as follows.

In case that the reaction is conducted by reacting a compound (II) orits reactive derivative at the amino group or a salt thereof with acompound (III) wherein A is a group of the formula: ##STR23##[hereinafter referred to as "oximino acylating agent" (III)] in thepresence of a condensing agent, for example, phosphorus pentachloride,thionyl chloride, etc., an anti isomer of the oximino compound (I) tendsto be produced as the dominant product and the corresponding syn isomerthereof can be hardly isolated from the reaction product even when a synisomer of the oximino acylating agent (III) is used. It may beunderstood that the tendency of such a isomerization in the reactionconducted by the method as explained above is due to the fact that theless stable syn isomer tends to isomerize partially or wholly to thecorresponding more stable anti isomer in the course of such reaction,for example, in so-called activation step of the oximino acylating agent(III) so that more stable isomer, i.e. the anti isomer of the oximinocompound (I) may be isolated as the reaction product.

Accordingly, in order to obtain a syn isomer of the oximino compound (I)selectively and in high yield, it is preferable to use a syn-isomer ofthe oximino acylating agent (III), and to conduct the reaction under aselected reaction condition. That is, a syn isomer of the oximinocompound (I) can be obtained selectively and in high yield by conductingthe reaction of a compound (II) with a syn isomer of the oximinoacylating agent (III), for example, in the presence of a Vilsmeierreagent as mentioned above and under around neutral condition.

The object compound (I) and salt thereof are useful as an antimicrobialagent, and a part thereof can be also used as a starting material in thefollowing processes.

Process B: C-Nitrosation

An object compound (Ib) and its salt can be prepared by reacting acompound (IV) or its salt with a nitrosating agent.

The starting compound (IV) corresponds to the 3-cephem compound (I)wherein R¹ is haloacetyl, R⁴ is hydrogen, halogen, lower alkyl or agroup of the formula: --O--R⁷ in which R⁷ is lower alkyl and A ismethylene, and can be prepared by the above Process A, preferably byreacting a compound (II) with diketene and halogen (e.g. chlorine,bromine, etc.). This prepared starting compound (IV) can be used in thisprocess without any isolation and/or purification.

Suitable nitrosating agent may include nitrous acid and its conventionalderivatives such as nitrosyl halide (e.g. nitrosyl chloride, nitrosylbromide, etc.), alkali metal nitrite (e.g. sodium nitrite, potassiumnitrite, etc.), alkyl nitrite (e.g. butyl nitrite, pentyl nitrite, etc.)and the like.

In case that a salt of nitrous acid is used as a nitrosating agent, thereaction is preferably carried out in the presence of an acid such as aninorganic or organic acid (e.g. hydrochloric acid, sulfuric acid, formicacid, acetic acid, etc.). And also, in case that an ester of nitrousacid is used, the reaction is preferably carried out in the presence ofa strong base such as alkali metal alkoxide or the like.

This reaction is usually conducted in a solvent such as water, aceticacid, benzene, methanol, ethanol, tetrahydrofuran or any other solventwhich does not adversely influence the reaction. The reactiontemperature is not critical and the reaction is preferably conductedwithin the range of cooling to an ambient temperature.

Thus prepared compound (Ib) and salt thereof can be used as a startingmaterial in the following Processes C and D.

Process C: Etherification

An object compound (Ic) and its salt can be prepared by reacting acompound (V) or its salt with an etherifying agent.

The starting compound (V) corresponds to the compound (I) wherein A isN-hydroxyiminomethylene group, and can be prepared by the above ProcessA and B and also by the following Process D.

Suitable examples of the etherifying agent may include a conventionalalkylating agent such as dialkyl sulfate (e.g. dimethyl sulfate, diethylsulfate, etc.), diazoalkane (e.g. diazomethane, diazoethane, etc.),alkyl halide (e.g. methyl iodide, ethyl iodide, ethyl bromide, etc.),alkyl sulfonate (e.g. methyl tosylate, etc.), the correspondingalkenylating-, alkynylating- or cycloalkylating agent, in which thealiphatic hydrocarbon moiety may be substituted with halogen, carboxy oresterified carboxy, for example, alkenyl halide (e.g. allyl iodide,etc.), alkynyl halide (e.g. propargyl bromide, etc.), cycloalkyl halide(e.g. cyclohexyl bromide, etc.), lower alkoxycarbonylalkyl halide (e.g.ethoxycarbonylmethyl iodide, etc.) and the like.

In case of using diazoalkane as an etherifying agent, the reaction isusually conducted in a solvent such as diethyl ether, dioxane or anyother solvent which does not adversely influence the reaction, at atemperature within a range of cooling to an ambient temperature.

In case of using the other etherifying agent, the reaction is usuallyconducted in a solvent such as water, acetone, ethanol, diethyl ether,dimethylformamide or any other solvent which does not adverselyinfluence the reaction within a temperature range of cooling to heating,preferably in the presence of a base such as an inorganic or organicbase, suitable examples of which are referred to the ones used for thebasic hydrolysis in the Process E as illustrated below.

Some of the object compound (Ic) and salt thereof are useful as anantimicrobial agent, and some of them, especially the compound where R¹is haloacetyl can be used as a starting material in the followingProcess D.

This process is an alternative one for preparing the compound (Ic) whereR¹ is haloacetyl group, and further this process is particularlypreferable and advantageous for preparing the compound (Ic) where R¹ ishaloacetyl and R_(a) ² is substituted- or unsubstituted-lower alkyl,lower alkenyl or lower alkynyl, more preferably lower alkyl.

Process D: Thiazole ring formation

A compound (Id) and its salt can be prepared by reacting a compound (VI)or its salt with a thiourea compound (VII).

The starting compound (VI) corresponds to the 3-cephem compound (I)wherein R¹ is haloacetyl, R⁴ is hydrogen, halogen, lower alkyl or agroup of the formula: --O--R⁷ in which R⁷ is lower alkyl and A is agroup of the formula: ##STR24## in which R² is as defined above, and canbe prepared by the above Process(es) A, B and/or C.

The reaction is usually conducted in a solvent such as water, alcohol(e.g. methanol, ethanol, etc.), benzene, dimethylformamide,tetrahydrofuran or any other solvent which does not adversely influencethe reaction within a temperature range of an ambient temperature toheating.

This process is an alternative and highly advantageous one for providingthe active compound (Id), especially (a) the compound (Id) wherein R² ishydrogen and R⁶ is amino from the compound (IV) via the Process B, and(b) the compound (Id) wherein R² is lower alkyl and R⁶ is amino from thecompound (IV) via the Processes B and C.

Process E: Elimination of amino-protective group

A compound (Ie) and its salt can be prepared by subjecting a compound(VIII) or its salt to elimination reaction of the protective group inthe protected amino group for R_(a) ⁶.

The starting compound (VIII) corresponds to the compound (I) wherein R¹is thiazolyl of the formula: ##STR25## in which R_(a) ⁶ is protectedamino and A is a group of the formula: ##STR26## in which R² is asdefined above, and can be prepared for example, by the above Process A.

The elimination reaction may be conducted in accordance with aconventional method such as hydrolysis, reduction or the like. Thesemethods may be selected according to the kind of the protective group tobe eliminated.

The hydrolysis may include a method using an acid (acidic hydrolysis), abase (basic hydrolysis) or hydrazine, and the like.

Among these methods, hydrolysis using an acid is one of the common andpreferable methods for eliminating the protective group such as an acylgroup, for example, substituted or unsubstituted lower alkanoyl,substituted or unsubstituted lower alkoxycarbonyl, substituted orunsubstituted ar(lower)alkoxycarbonyl, lower cycloalkoxycarbonyl,substituted phenylthio, substituted alkylidene, substitutedaralkylidene, substituted cycloalkylidene or the like, particulars ofwhich are to be referred to those as illustrated for the N-protectivegroup, respectively.

Suitable acid to be used in this acidic hydrolysis may include anorganic or inorganic acid such as formic acid, trifluoroacetic acid,benzenesulfonic acid, p-toluenesulfonic acid, hydrochloric acid,cation-exchange resin, and the like. Preferable acid is the one whichcan be easily separated out from the reaction product by a conventionalmanner such as neutralization or distillation under reduced pressure,for example, formic acid, trifluoroacetic acid, hydrochloric acid or thelike. The acid suitable for the reaction can be selected inconsideration of the chemical property of the starting compound and theproduct as well as the kind of the protective group to be eliminated.The acidic hydrolysis can be conducted in the presence or absence of asolvent. Suitable solvent may be a conventional organic solvent, wateror a mixture thereof, which does not adversely influence this reaction.Particularly, when the hydrolysis is conducted with trifluoroaceticacid, the reaction may be accelerated by addition of anisole.

The hydrolysis using a base can be applied for eliminating theprotective group such as an acyl group, preferably, for example,haloalkanoyl (e.g. trifluoroacetyl, etc.) and the like. Suitable basemay include, for example, an inorganic base such as alkali metalhydroxide (e.g. sodium hyroxide, potassium hydroxide, etc.), alkalineearth metal hydroxide (e.g. magnesium hydroxide, calcium hydroxide,etc.), alkali metal carbonate (e.g. sodium carbonate, potassiumcarbonate, etc.), alkaline earth metal carbonate (e.g. magnesiumcarbonate, calcium carbonate, etc.), alkali metal bicarbonate (e.g.sodium bicarbonate, potassium bicarbonate, etc.), alkaline earth metalphosphate (e.g. magnesium phosphate, calcium phosphate, etc.), alkalimetal hydrogen phosphate (e.g. disodium hydrogen phosphate, dipotassiumhydrogen phosphate, etc.), or the like, and an organic base such asalkali metal acetate (e.g. sodium acetate, potassium acetate, etc.),trialkylamine (e.g. trimethylamine, triethylamine, etc.), picoline,N-methylpyrrolidine, N-methylmorpholine,1,5-diazabicyclo[4,3,0]-5-nonene, 1,4-diazabicyclo[2,2,2]octane,1,5-diazabicyclo[5,4,0]-7-undecene, anion-exchange resin or the like.The hydrolysis using a base is often carried out in water or aconventional organic solvent or a mixture thereof.

The hydrolysis using hydrazine can be applied for eliminating theprotective group such as dibasic acyl, for example, succinyl, phthaloylor the like.

The reduction can be applied for eliminating the protective group suchas acyl, for example, halo(lower)alkoxycarbonyl (e.g.trichloroethoxycarbonyl, etc.), substituted or unsubstitutedar(lower)alkoxycarbonyl (e.g. benzyloxycarbonyl,p-nitrobenzyloxycarbonyl, etc.), 2-pyridylmethoxycarbonyl, etc., aralkyl(e.g. benzyl, benzhydryl, trityl, etc.) and the like. Suitable reductionmay include, for example, reduction using an alkali metal borohydride(e.g. sodium borohydride, etc.), conventional catalytic hydrogenolysisand the like.

And further, the protective group such as halo(lower)alkoxycarbonyl or8-quinolyloxycarbonyl can be eliminated by treatment with a heavy metalsuch as copper, zinc or the like.

The reaction temperature is not critical and may be optionally selectedin consideration of the chemical property of the starting compound andreaction product as well as the kind of the N-protective group and themethod to be applied, and the reaction is preferably carried out under amild condition such as under cooling, at ambient temperature or slightlyelevated temperature.

The process includes in its scope the cases that the functionallymodified carboxy for R⁵ is simultaneously transformed into the freecarboxy group in the course of the above reaction or in thepost-treatment.

As to this process, it is to be understood that the purpose of thisprocess lies in providing the generally more active compound (I')wherein R_(a) ¹ is aminothiazolyl by eliminating the protective group inthe protected amino group of the compound (VIII) prepared by the otherprocesses as mentioned above or below.

Process F: Reductive formation of 3-hydroxycepham

A compound (If) and its salt can be prepared by reducing a compound (IX)or its salt.

The starting compound (IX) corresponds to the 3-cephem compound (I)wherein R¹ is thiadiazolyl or thiazolyl of the formula: ##STR27## inwhich R⁶ is as defined above, R³ is hydrogen, R⁴ is a group of theformula: --O--R⁷ in which R⁷ is hydrogen and A is a group of theformula: ##STR28## in which R² is as defined above, and can be prepared,for example, by the above Process A.

The method of reduction applied to this process may include aconventional one which is applicable for reduction of ketonic carbonylgroup including its tautomeric enol form into hydroxymethylene group,and the preferable method may be reduction using an alkali metalborohydride (e.g. sodium borohydride, etc.) or a combination of an acid(e.g. hydrochloric acid, sulfuric acid, formic acid, acetic acid, etc.)and a metal (e.g. zinc, iron, copper, etc.), catalytic reduction using aconventional catalyst (e.g. palladium on carbon, palladium sponge, Raneynickel, platinum, platinum black, etc.) or the like.

The reaction is usually carried out in a conventional solvent such aswater, alcohol (e.g. methanol, ethanol, etc.), dimethylformamide,tetrahydrofuran or any other solvent which does not adversely influencethe reaction within a temperature range from cooling to somewhatelevated temperature.

Although thus prepared compound (If) and salt thereof have antimicrobialactivities, they are also useful mainly as an intermediate, especiallyas a starting material in the following Process G and successivelyProcess H for preparing the more active 3-cephem compound (Ih).

Process G: O-acylation

A compound (Ig) and its salt can be prepared by reacting a compound (If)or its salt with a compound (X), its salt or its reactive derivative.

As to the compound (X), suitable examples of the acyl moiety for R_(a) ⁷are to be referred to those as exemplified above for the acyl group forR⁷ of the compound (I).

The reactive derivative of the compound (X) may be an acyl halide,anhydride, azide, activated ester, activated amide and the like, whichare to be referred to those as exemplified above for the compound (III)in the Process A, preferably an acyl halide such as lower alkanoylhalide (e.g. acetyl chloride, etc.), aroyl halide (e.g. benzoylchloride, etc.), lower alkanesulfonyl halide (e.g. mesyl chloride, mesylbromide, ethanesulfonyl chloride, etc.), arenesulfonyl halide (e.g.tosyl chloride, etc.), an acyl azide such as lower alkanesulfonyl azide(e.g. mesyl azide, etc.), arenesulfonyl azide (e.g. tosyl azide, etc.)or the like, and more preferably lower alkanesulfonyl halide orarenesulfonyl halide.

The reaction is usually carried out in a conventional solvent such asdimethylformamide, chloroform, methylene chloride or any other solventwhich does not adversely influence the reaction, under cooling or at anambient or somewhat elevated temperature.

In case that the acyl halide is used as an acylating agent, the reactionis generally conducted in the presence of a base as exemplified in theabove Process E.

This process is the first activation step for preparing a more active3-cephem compound (Ih) from the 3-hydroxycepham compound (If) via the3-acyloxycepham compound (Ig), which is successively treated with a basein the following Process H.

Process H: 3-Cephem formation

This process is the final step to transform the 3-hydroxycephem compound(IX) into the more active 3-cephem compound (Ih) or its salt. That is, acompound (Ih) or its salt can be prepared by treating a compound (Ig) asprepared in the above Process G or its salt with a base.

The preferable base includes an inorganic base such as metal hydroxide(e.g. sodium hydroxide, potassium hydroxide, etc.), metal carbonate(e.g. sodium carbonate, potassium carbonate, magnesium carbonate, etc.),metal bicarbonate (e.g. sodium bicarbonate, potassium bicarbonate,etc.), organic base such as tertiary amine (e.g. trimethyl amine,triethyl amine, pyridine, etc.) alkali metal alkoxide (e.g. sodiummethoxide, sodium ethoxide, etc.) and the like.

The reaction is usually carried out in a conventional solvent such as analcohol, dimethylformamide, chloroform, methylene chloride or any othersolvent which does not adversely influence the reaction, under coolingor at an ambient or somewhat elevated temperature.

Process I: Halogenation

A compound (Ii) or its salt can be prepared by halogenating a compound(XI) or its salt.

The starting compound (XI) corresponds to the compound (I) wherein R¹ isthiadiazolyl or thiazolyl of the formula: ##STR29## in which R⁶ is asdefined above, R³ is hydrogen, R⁴ is a group of the formula: --O--R⁷ inwhich R⁷ is hydrogen and A is a group of the formula: ##STR30## in whichR² is as defined above, and can be prepared by the processes asexplained above.

Suitable halogenating agent may include a conventional halogen compoundsuch as phosphorus halide (e.g. phosphorus trichloride, phosphoruspentachloride, phosphorus tribromide, phosphorus pentabromide,phosphoryl chloride, etc.), thionyl chloride and the like.

The reaction is usually carried out in a conventional solvent such aschloroform, methylene chloride, dimethylformamide or any other solventwhich does not adversely influence the reaction and preferably undercooling or at ambient or somewhat elevated temperature.

Process J: Esterification

This process is to provide an ester compound (Ij) and its salt forimproving the chemical, phisiological and/or pharmaceutical propertiesof the corresponding free carboxy compound (XII), which corresponds tothe 3-cephem compound (I) wherein R⁵ is carboxy, or its salt.

The esterification is conducted by reacting a free carboxy compound(XII), its reactive derivative at the carboxy or a salt thereof with anesterifying agent.

The preferred reactive derivative at the carboxy group of the compound(XII) is to be referred to those of the compound (III) as exemplified inthe Process A.

The esterifying agent may include a hydroxy compound and its reactionequivalent.

Suitable examples of the hydroxy compound may be a substituted orunsubstituted alcohol such as alkanol, aralkanol, arenol or the like,particulars of which may be substituted alcohol such asalkanoyloxy(lower)alkanol (e.g. acetoxymethanol, propionyloxymethanol,butyryloxymethanol, pentanoyloxymethanol, hexanoyloxymethanol,acetoxyethanol, propionyloxyethanol, butyryloxyethanol,pentanoyloxyethanol, hexanoyloxyethanol, acetoxypropanol,propionyloxypropanol, hexanoyloxypropanol, hexanoyloxyhexanol,palmitoyloxymethanol, etc.), halo(lower)alkanol (e.g. mono-, di- ortrichloroethanol, etc.), lower cycloalkyl(lower)alkanol (e.g.1-cyclopropylethanol, etc.), substituted ar(lower)alkanol (e.g.4-nitrobenzyl alcohol, 4-chlorobenzyl alcohol, 4-methoxybenzyl alcohol,3,5-di-tert-butyl-4-hydroxybenzyl alcohol, bis(methoxyphenyl)methanol,etc.), substituted arenol (e.g. 4-methoxyphenol, etc.), thecorresponding unsubstituted alcohol or the like.

Suitable reactive equivalent of the hydroxy compound may include aconventional one such as halide, alkanesulfonate, arenesulfonate or saltof the hydroxy compound, diazoalkane, diazoaralkane, and the like.

Preferable halide of the hydroxy compound may be chloride, bromide oriodide.

Preferable alkane- or arene-sulfonate of the hydroxy compound may bemethanesulfonate, ethanesulfonate, benzenesulfonate, tosylate or thelike.

Preferable salt of the hydroxy compound may be an alkali metal salt suchas lithium salt, sodium salt, potassium salt or the like.

Preferable diazoalkane and diazoaralkane may be diazomethane,diazoethane, diazopropane, diphenyldiazomethane or the like.

The reaction can be carried out in the presence or absence of a solventsuch as N,N-dimethylformamide, dimethylsulfoxide or any other solventwhich does not adversely influence the reaction, and within atemperature range of cooling to heating. The liquid hydroxy compound canbe also used as a solvent in this reaction.

This reaction can be preferably conducted in the presence of aninorganic or organic base as exemplified in the above Process E.

In case of preparing a substituted- or unsubstituted-aryl ester (Ij),particularly substituted- or unsubstituted-phenyl ester, this reactionis to be conducted by reacting (a) a compound (XII) or its salt withphenol or its salt in the presence of a condensing agent as exemplifiedin the above Process A, or (b) a reactive derivative of the compound(XII) preferably a mixed acid anhydride of the compound (XII) withphenol or its salt in the presence of a base.

In case that a compound (XII), where A is a group of the formula:##STR31## in which R² is an aliphatic hydrocarbon residue substitutedwith carboxy, is used as a starting material in this reaction, the saidcarboxy group may be also esterified in accordance with the reagent andthe reaction conditions, and this mode of the reaction is includedwithin the scope of this process.

And further, in case that the 2-cephem compound corresponding to thecompound (Ij) is produced, the said 2-cephem compound can be transformedinto the 3-cephem compound (Ij) by oxydizing and then reducing theresultant S-oxide compound in a conventional manner. This mode of thereactions is also included within the scope of this process.

Process K: Carboxy formation

This process is to provide a free carboxy compound (Ik) or its salt,especially the compound (Ik) wherein R¹ is thiadiazolyl or thiazolyl ofthe formula: ##STR32## in which R⁶ is as defined above and A is a groupof the formula: ##STR33## in which R² is as defined above, whichgenerally exhibits higher antimicrobial activities as compared with thecorresponding functionally modified carboxy compound (XIII).

Accordingly, the meaning of the functionally modified carboxy in thecompound (XIII) lies in mainly synthetic manufacture by chemicalprocess(es) as illustrated hereinabove.

This process is conducted by transforming the functionally modifiedcarboxy group of the starting compound (XIII) into free carboxy group,and the preferred functionally modified carboxy for R_(b) ⁵ in thecompound (XIII) may be an esterified carboxy group as exemplified for R⁵of the compound (I).

The method to be applied to this process includes conventional ones suchas hydrolysis, reduction and the like.

The method of hydrolysis includes a conventional one using an acid,base, enzyme or enzymatic preparation, and the like.

Suitable examples of the acid and base are to be referred to those asexemplified in the above Process E, and the acidic or basic hydrolysiscan be carried out in a similar manner to that of the Process E.

Suitable enzyme includes an esterase and esterase preparation whichexhibits an esterase activity such as a cultured broth of microorganismor processed materials of microorganism, the preparation of animal orplant tissues, or the like, and preferably a cultured broth ofmicroorganism or processed material thereof.

An esterase to be used in the enzymatic hydrolysis may be used not onlyin a purified state, but also in a crude state.

Such an esterase is frequently found to exist widely, for example, invarious kind of microorganisms, which can be easily isolated from a soilsample and other sources by conventional means, and further can beeasily selected from the collected cultures available in publicfacilities for culture collection such as ATCC (American Type CultureCollection, Maryland, USA), IAM (Institute of Applied Microbiology,University of Tokyo, Japan), IFO (Institute For Fermentation, Osaka,Japan), IID (The Institute for Infectious Diseases, University of Tokyo,Tokyo, Japan), CBS (Centraalbureau voor Schimmelcultures, Bearn,Netherlands), FERM (Fermentation Research Institute, Agency ofIndustrial Science and Technology, Chiba, Japan) and NRRL (NorthernUtilization Research and Development Division, U.S. Department ofAgriculture, Illinois, U.S.A.) and the like.

As to the microorganism having an esterase activity, there may beexemplified one belonging to the genus, Bacillus, Corynebacterium,Micrococcus, Flavovacterium, Salmonella, Staphylococcus, Vibrio,Microbacterium, Escherichia, Arthrobacter, Azotobacter, Alcaligenes,Rhizobium, Brevibacterium, Kluyvera, Proteus, Sarcina, Pseudomonas,Xanthomonas, Protaminobacter, Comamonus and the like.

Examples of the above microorganisms may be Bacillus subtilis IAM-1069,IAM-1107, IAM-1214, Bacillus sphaericus IAM-1286, Corynebacterium equiIAM-1308, Micrococcus varians IAM-1314, Flavobacterium rigeus IAM-1238,Salmonella typhimurium IAM-1406, Staphylococcus epidermidis IAM-1296,Microbacterium flavum IAM-1642, Alcaligenes faecalis ATCC-8750,Arthrobacter simplex ATCC-6946, Azotobacter vinelandii IAM-1078,Escherichia coli IAM-1101, Rhizobium japonicum IAM-0001, Vibriometchnikovii IAM-1039, Brevibacterium helvolum IAM-1637, Protaminobacteralboflavum IAM-1040, Comamonas terrigena IFO-12685, Sarcina luteaIAM-1099, Pseudomonus schuylkillienis IAM-1055, Xanthomonas trifoliiATCC-12287 or the like.

In the enzymatic hydrolysis, the esterase can be preferably used in aform of a cultured broth obtained by culturing microorganisms having anesterase activity in a suitable manner, or of its processed material.

Cultivation of microorganisms can be generally conducted in aconventional manner. As a culture medium to be used, there may be used anutrient one containing sources of assimilable carbon and nitrogen andinorganic salts. The preferred sources of carbon are, for example,glucose, sucrose, lactose, sugars, glycerol and starch. The preferredsources of nitrogen are, for example, meat extract, peptone, glutenmeal, corn meal, cotton-seed meal, soybean meal, corn steep liquor,yeast extracts, casein hydrolysate and amino acids, as well as inorganicand organic nitrogen such as ammonium salts (e.g. ammonium sulfate,ammonium nitrate, ammonium phosphate, etc.), sodium nitrate or the like.If desired, mineral salts such as calcium carbonate, sodium or potassiumphosphate, magnesium salts and copper salts, and various vitamines canbe also used.

Suitable pH of the culture medium, suitable cultivation temperature andsuitable cultivation time vary with the kind of the microorganisms to beused. A desirable pH usually lies in a range of pH 5 to 8. Thetemperature is usually selected from about 20° C. to about 35° C. Thecultivation time is usually selected from 20 hours to 120 hours.

The cultured broth per se thus obtained and its processed material maybe employed for enzymatic hydrolysis of this process. The "processedmaterial" of cultured broth means any preparation having esteraseactivity, which is processed by conventionally suitable means forincreasing said esterase activity.

The esterase activity of the cultured broth is present in cells(intracellularly) and/or out of cells (extracellularly).

When the activity exists mainly in cells, the following preparation, forexample, may be used as a processed material of the cultured broth.

(1) raw cells; separated from the cultured broth in conventional mannerssuch as filtration and centrifugation,

(2) dried cells; obtained by drying said raw cells in conventionalmanners such as lyophilization and vacuum drying,

(3) a cell-free extract; obtained by destroying said raw or dried cellsin conventional manners (e.g. grinding the cells with almina, sea sand,etc. or treating the cells with super sonic waves), or

(4) an enzyme solution; obtained by purification or partial purificationof said cell-free extract in a conventional manner.

When the activity exists mainly out of cells, the following preparation,for example, may be used as a processed material.

(1) a supernatant or a filtrate; obtained from the cultured broth in aconventional manner, or

(2) an enzyme solution; obtained by purification or partial purificationof said supernatant or filtrate in a conventional manner.

The enzymatic hydrolysis is conducted by contacting the compound (XIII)with the cultured broth of the microorganism or its processed materialin an aqueous medium such as water or a buffer solution (e.g. phosphatebuffer, etc.), preferably in the presence of conventional surface-activeagent. That is, the reaction is usually conducted by adding the compound(XIII) to the cultured broth of the microorganism or its liquidprocessed material (e.g. supernatant, filtrate, enzyme solution, etc.),or to the solution or suspension of the cultured broth or its processedmaterial in an aqueous medium. Sometimes, an agitation of the saidreaction mixture is preferable.

Preferred pH of reaction mixture, concentration of substrates, reactiontime and reaction temperature may vary with characteristics of thecultured broth or its processed material to be used, or the compound(XIII) to be used. However, the reaction conditions are preferablyselected from a range of at pH 4 to 10, more preferably at pH 6 to 8, at20° to 50° C., more preferably at 25° to 35° C. for 1 to 100 hours. Theconcentration of the starting compound (XIII) to be used as a substratein the reaction mixture may be in a range of 0.1 to 100 mg per ml,preferably 1 to 20 mg per ml.

The method of the reduction for this process may be carried out in asimilar manner to that of the above Process E.

This process includes within its scope the cases that the protectivegroup in the protected amino for R⁶, which is a substituent on thethiazolyl group for R¹, is eliminated and/or the esterified carboxygroup, which is an optional substituent on the aliphatic hydrocarbonresidue for R² in the group A, is transformed into free carboxy group inthe course of the reaction or the post-treatment.

The compound obtained in accordance with the processes as explainedabove can be isolated and purified in a conventional manner.

In case that the object compound (I) has free carboxy for R⁵ and/or freeamino for R⁶, it may be transformed into its pharmaceutically acceptablesalt by a conventional method.

Among the object compound (I), the compound (I'), its pharmaceuticallyacceptable salt and bioprecursor thereof exhibit high antimicrobialactivities inhibiting the growth of a wide variety of pathogenicmicroorganisms including Gram-positive and Gram-negative bacteria andare useful as antimicrobial agents.

And further, the compound (I") and its salt are novel and useful as anintermediate for preparing the active compound (I'), itspharmaceutically acceptable salt or bioprecursor thereof.

According to the aforementioned processes, more specifically thefollowing compounds can be prepared.

7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-2,3-dimethyl-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (anti isomer)

7-[2-(1,2,3-thiadiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-tosyloxy-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-isopropoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-propoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-isobutyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-n-butoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-n-hexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-allyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-propargyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-carboxymethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-n-pentyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-(2-chloroethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (anti isomer)

7-[2-(2-amino-4-thiazolyl)-2-n-octyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-(2,3,3-trifluoro-2-propenyloxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-lauroyloxymethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(1,2,3-thiadiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-n-butoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-n-butoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-propargyloxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-trifluoromethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

the corresponding functionally modified derivative such as

hexanoyloxymethyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

pivaloyloxymethyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer)

4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer)

4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-n-propoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer)

4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-isobutoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer)

the corresponding salt such as

sodium7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

calcium7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

magnesium7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

arginine salt of 7-[2-(2-amino-4-thiazolyl)-2methoxyiminoacetamido]-3-cephem-4-carboxylic acid (syn isomer)

lysine salt of7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid hydrochloride (syn isomer)

In order to show the utility of the active compound (I'), the test dataof some representative compounds (I') are shown in the following.

1. In vitro antibacterial activity (1) Test method

In vitro antibacterial activity was determined by the two-foldagar-plate dilution method as described below.

One loopful of the 100-fold dilution of an overnight culture of eachtest strain in Trypticase-soy broth was streaked on heart infusion agar(HI-agar) containing graded concentrations of the test compound andincubated at 37° C. for 20 hours. The minimal inhibitory concentration(MIC) was expressed in μg/ml.

    ______________________________________                                        (2) Test compounds:                                                           No.                                                                           ______________________________________                                        1   7-[2-(2-Amino-4-thiazolyl)-2-methoxyiminoacetamido]-                          3-cephem-4-carboxylic acid (syn isomer)                                   2   7-[2-(2-Amino-4-thiazolyl)-2-hydroxyiminoacetamido]-                          3-cephem-4-carboxylic acid (syn isomer)                                   3   7-[2-(2-Amino-4-thiazolyl)-2-ethoxyiminoacetamido]-                           3-cephem-4-carboxylic acid (syn isomer)                                   4   7-[2-(2-Amino-4-thiazolyl)-2-methoxyiminoacetamido]-                          3-chloro-3-cephem-4-carboxylic acid (syn isomer)                          5   7-[2-(2-Amino-4-thiazolyl)-2-n-propoxyiminoacetamido]-                        3-cephem-4-carboxylic acid (syn isomer)                                   6   7-[2-(2-Amino-4-thiazolyl)-2-n-butoxyiminoacetamido]-                         3-cephem-4-carboxylic acid (syn isomer)                                   7   7-[2-(2-Amino-4-thiazolyl)-2-allyloxyiminoacetamido]-                         3-cephem-4-carboxylic acid (syn isomer)                                   8   7-[2-(2-Amino-4-thiazolyl)-2-propargyloxyiminoacetamido]-                     3-cephem-4-carboxylic acid (syn isomer)                                   9   7-[2-(2-Amino-4-thiazolyl)-2-n-pentyloxyiminoacetamido]-                      3-cephem-4-carboxylic acid (syn isomer)                                   10  7-[2-(2-Amino-4-thiazolyl)-2-n-hexyloxyiminoacetamido]-                       3-cephem-4-carboxylic acid (syn isomer)                                   11  7-[2-(2-Amino-4-thiazolyl)-2-cyclohexyloxyiminoacetamido]-                    3-cephem-4-carboxylic acid (syn isomer)                                   12  7-[2-(2-Amino-4-thiazolyl)-2-(2-chloroethoxyimino)-                           acetamido]-3-cephem-4-carboxylic acid (syn isomer)                        13  7-[2-(2-Amino-4-thiazolyl)-2-(2,2,2-trifluoroethoxy-                          imino)acetamido]-3-cephem-4-carboxylic acid (syn                              isomer)                                                                   ______________________________________                                    

    __________________________________________________________________________    (3) Test results:                                                             MIC (μg/ml)                                                                         Compound No.                                                         Test Strains                                                                           1    2    3   4   5   6   7   8   9   10  11  12  13                 __________________________________________________________________________    Staphylococcus                                                                         6.25 0.39 3.13                                                                              12.5                                                                              1.56                                                                              0.78                                                                              1.56                                                                              1.56                                                                              0.39                                                                              1.56                                                                              0.39                                                                              1.56                                                                              1.56               aureus 209P JC-1                                                              Escherichia                                                                            ≦0.025                                                                      0.1  0.05                                                                              0.39                                                                              0.2 0.39                                                                              0.2 0.1 3.13                                                                              1.56                                                                              3.13                                                                              0.1 0.2                coli NIHJ JC-2                                                                Proteus  ≦0.025                                                                      0.1  ≦0.025                                                                     ≦0.025                                                                     ≦0.025                                                                     0.05                                                                              ≦0.025                                                                     ≦0.025                                                                     0.39                                                                              0.2 0.78                                                                              ≦0.025                                                                     ≦0.025      vulgaris IAM-1025                                                             Klebsiella                                                                             ≦0.025                                                                      ≦0.025                                                                      ≦0.025                                                                     0.1 ≦0.025                                                                     0.2 ≦0.025                                                                     ≦0.025                                                                     0.2 0.05                                                                              0.39                                                                              0.1 0.05               pneumoniae 20                                                                 Proteus  ≦0.025                                                                      ≦0.025                                                                      ≦0.025                                                                     0.0125                                                                            0.1 0.2 ≦0.025                                                                     ≦0.025                                                                     1.56                                                                              0.78                                                                              1.56                                                                              0.2 0.2                mirabilis 18                                                                  Pseudomonus                                                                            0.39 6.25 ≦1.56                                                                      6.25                                                                              ≦1.56                                                                      ≦1.56                                                                      ≦1.56                                                                      ≦1.56                                                                      3.13                                                                              ≦1.56                                                                      ≦1.56                                                                      ≦1.56                                                                      ≦ 1.56      aeruginosa                                                                    NCTC-10490                                                                    Serratia 1.56 12.5 0.78                                                                              50  3.13                                                                              6.25                                                                              1.56                                                                              3.13                                                                              3.13                                                                              12.5                                                                              12.5                                                                              12.5                                                                              6.25               marcescens 35                                                                 __________________________________________________________________________

2. Protecting effect against experimental infections in mice (1) Testmethod

Male ICR strain mice aged 4 weeks, each weighing 18.5-21.5 g. were usedin groups of 10 mice. The test bacteria were cultured overnight at 37°C. on Trypticase-soy agar and then suspended in 5% mucin to obtain thesuspension corresponding to each challenge cells. Mice were inoculatedintraperitoneally with 0.5 ml. of the suspension. A solution containingeach test compound was given subcutaneously to the mice in variousdosage one hour after the challenge. The ED₅₀ values were calculatedfrom the number of surviving mice for each dosage after four days ofobservation.

    ______________________________________                                        (2) Test compounds                                                            No.                                                                           ______________________________________                                        1      7-[2-(2-Amino-4-thiazolyl)-2-methoxyiminoacetamido]-                          3-cephem-4-carboxylic acid (syn isomer)                                reference                                                                            7-[2-(2-Amino-4-thiazolyl)-2-methoxyimino-                                    acetamido]cephalospolanic acid (syn isomer)                            ______________________________________                                    

    __________________________________________________________________________    (3) Test results:                                                                            ED.sub.50 (s.c.)(mg/kg)                                                                 MIC (μg/ml.)                                      Test    Inoculated                                                                           Test Compounds                                                                          Inoculum                                                                           Test Compounds                                  Bacteria                                                                              Cells/mouse                                                                          1    reference                                                                          Size 1   reference                                   __________________________________________________________________________    Escherichia                                                                           1.1 × 10.sup.7                                                                 0.95 2.8  10.sup.0 *.sup.1                                                                   0.78                                                                              3.13                                        coli 54                  10.sup.-2 *.sup.2                                                                  0.05                                                                              0.1                                         Klebsiella                                                                              8 × 10.sup.6                                                                 <0.98                                                                              0.995                                                                              10.sup.0                                                                           0.39                                                                              3.13                                        pneumoniae 39            10.sup.-2                                                                          ≦0.025                                                                     0.05                                        Proteus 9.9 × 10.sup.6                                                                 0.39 1.171                                                                              10.sup.0                                                                           1.56                                                                              50                                          rettgeri 24              10.sup.-2                                                                          ≦0.025                                                                     0.1                                         Serratia                                                                              1.2 × 10.sup.7                                                                 3.562*.sup.3                                                                       31.427*.sup.3                                                                      10.sup.0                                                                           25  50                                          Marcescens 58            10.sup.-2                                                                          0.39                                                                              1.56                                        __________________________________________________________________________     *.sup.1 overnight culture                                                     *.sup.2 100fold dilution of the overnight culture                             *.sup.3 treated with two divisional doses at 1 hr. and 3 hrs. after           infection                                                                

3. Acute toxicity (1) Test method

Ten male and 10 female rats aged 6 weeks (JCL-SD strain) were used pergroup. Test compound dissolved in distilled water was givensubcutaneously and intraveneously to the animals. These animals wereobserved for 7 days after dosing. The LD₅₀ values were calculated fromthe number of dead animals by the Litchfield-Wilcoxon method.

(2) Test compound

7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

    ______________________________________                                        (3) Test results:                                                             Test                  LD.sub.50 (mg./kg.)                                     animal  Sex           s.c.    i.v.                                            ______________________________________                                        Rat     Male          >8000   about 8000                                              Female        >8000   >8000                                           ______________________________________                                    

4. Absorbability (1) Test method

Test compound was given orally to a group of 5 rats (JCL-SD strain,6-week-old, male) which had been fasted. Bile and urine samples werecollected at 0˜6 and 6˜24 hrs. The concentrations of the test compoundin the samples were determined by bioassay (disk method) using Batillussubtilis ATCC-6633 as test organism, and the recoveries in bile andurine were calculated.

(2) Test compound

7-[2-(2-amino-4-thiazolyl)-2-n-pentyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer)

(3) Test result:

Total recovery in bile and urine in 24 hrs. was 22.8%.

For prophylactic and/or therapeutic administration, the active compound(I') of the present invention is used in the form of conventionalpharmaceutical preparation which contains said compound, as an activeingredient, in admixture with pharmaceutically acceptable carriers suchas an organic or inorganic solid or liquid excipient which is suitablefor oral, parenteral or external administration. The pharmaceuticalpreparations may be in solid form such as capsule, tablet, dragee,ointment or suppository, or in liquid form such as solution, suspension,or emulsion. If needed, there may be included in the above preparationsauxiliary substances, stabilizing agents, wetting or emulsifying agents,buffers and the other commonly used additives.

While the dosage of the compounds may vary from and also depend upon theage and conditions of the patient, a kind of disease and a degree of theinfection, and further a kind of the active compound (I') to be applied,etc., an average single dose of about 50 mg., 100 mg., 250 mg. and 500mg. of the active compound (I') is sufficient for treating infectiousdiseases caused by pathogenic bacteria. In general, the active compound(I') can be administered in an amount between 1 mg/kg and 100 mg/kg,preferably 5 mg/kg and 50 mg/kg.

The starting compound (III) can be prepared as illustrated below.##STR34## wherein R_(a) ² is an aliphatic hydrocarbon residue which maybe substituted with halogen, carboxy or esterified carboxy

R_(a) ⁶ is protected amino

X is halogen

Y is lower alkoxycarbonyl, and

Z is lower alkyl

Each of the above processes are explained in the following.

Process 1: Etherification

The compound (III_(b)) and (III_(d)) can be prepared by reacting acompound (III_(a)) or (III_(c)) with an etherifying agent, respectively.

This reaction may be conducted substantially in the same manner as theaforementioned Process C.

Process 2: Thiazole ring formation

The compound (III_(c)) and (III_(d)) can be prepared by reacting acompound (III_(a)) or (III_(b)) with a thiourea compound (VII_(a)),respectively, and further the compound (III_(e)) can be prepared byreacting a compound (III_(b)) with thiourea.

This reaction may be conducted substantially in the same manner as theaforementioned Process D.

Process 3: Elimination of amino-protective group

The compound (III_(e)) and (III_(g)) can be prepared by subjecting acompound (III_(d)) or (III_(f)) to elimination reaction of theprotective group in the protected amino group for R_(a) ⁶, respectively.

This reaction may be conducted substantially in the same manner as theaforementioned Process E.

Process 4: Carboxy formation

The compound (III_(f)), (III_(g)) and (III_(j)) can be prepared bytransforming the esterified carboxy group of a compound (III_(d)),(III_(e)) or (III_(i)) into free carboxy group, respectively.

This reaction may be conducted substantially in the same manner as theaforementioned Process K.

Process 5: Oximation

The compound (I_(f)) can be also prepared by reacting a compound(III_(h)) with a hydroxylamine derivative (XIV) or its salt.

The hydroxylamine derivative (XIV) may be hydroxylamine substituted withan aliphatic hydrocarbon residue which may be substituted with halogen,carboxy or esterified carboxy, particulars of which are to be referredto those as exemplified before. Suitable salt of the hydroxylaminederivative (XIV) may be hydrochloride, hydrobromide, sulfate or thelike.

The reaction is usually conducted in a conventional solvent such aswater, alcohl, tetrahydrofuran, acetonitrile, dimethylsulfoxide,pyridine or any other solvent which does not adversely influence thereaction, or a mixture thereof, and the reaction temperature is notcritical.

In case that a salt of the hydroxylamine derivative (XIV) is used as areagent, the reaction is preferably conducted in the presence of aconventional base.

Process 6: Thiadiazol ring formation

The compound (III_(i)) can be prepared by reacting a compound (XV) witha hydrazine derivative (XVI), and then reacting the resultant compound(XVII) with sulfur dihalide (XVIII).

Among the starting compound (III), the compound of the formula:##STR35## wherein R_(a) ¹ is thiadiazolyl or thiazolyl of the formula:##STR36## in which R⁶ is amino or protected amino, R_(a) ^(2') is alkyl,alkenyl or alkynyl having more than one carbon atom or cycloalkyl whichmay be substituted with halogen, carboxy or esterified carboxy,

R⁸ is hydrogen or lower alkyl,

provided that R⁶ is amino which may be protected with formyl, and R⁸ ishydrogen, when R_(a) ^(2') is ethyl, isopropyl or allyl, is novel anduseful as a starting material in the aforementioned Process A.

Particulars of each definition in the above are to be referred to thoseas explained before.

Following examples are given only for explaining this invention in moredetail.

PREPARATION OF THE STARTING COMPOUNDS EXAMPLE A

(1) A solution of ethyl 2-methoxyiminoacetoacetate (a mixture of syn andanti isomers) (34.6 g.) and t-butoxycarbonylhydrazine (26.4 g.) inethanol (200 ml.) was stirred for 7.5 hours at ambient temperature andallowed to stand overnight to precipitate crystals. The crystals werecollected by filtration, washed with ethanol and dried to give ethyl2-methoxyimino-3-t-butoxycarbonylhydrazonobutyrate (a mixture of syn andanti isomers) (41.7 g.), mp 144° to 145° C.

I.R. ν_(max) ^(Nujol) : 3200, 1750, 1705, 1600, 1520 cm⁻¹.

N.M.R. δ_(ppm) (CDCl₃): 8.52 (1H, broad s), 4.35 (2H, q, J=7 Hz), 4.10(3H, s), 2.00 (3H, s), 1.50 (9H, s), 1.33 (3H, t, J=7 Hz).

(2) Sulfur dichloride (15.9 ml.) was added with stirring at ambienttemperature to a solution of ethyl2-methoxyimino-3-t-butoxycarbonylhydrazonobutyrate (a mixture of syn andanti isomers) (14.36 g.) in methylene chloride (150 ml.), and themixture was stirred for 1 hour at ambient temperature. To the reactionmixture was added ice-water (300 ml.), and the methylene chloride layerwas washed with water, with a saturated aqueous solution of sodiumbicarbonate and with a saturated aqueous solution of sodium chloride anddried over magnesium sulfate. The solvent was distilled off to give anoil. The oil was purified by column chromatography on silica gel using amixture of benzene and n-hexane (19:1) as an eluent to firstly giveethyl 2-methoxyimino-2-(1,2,3-thiadiazol-4-yl)acetate (syn isomer) (1.8g.), mp 77° to 79° C.

I.R. ν_(max) ^(Nujol) : 1720, 1595 cm⁻¹.

N.M.R. δ_(ppm) (CDCl₃): 8.92 (1H, s), 4.46 (2H, q, J=7 Hz), 4.06 (3H,s), 1.38 (3H, t, J=7 Hz).

From subsequent fractions, ethyl2-methoxyimino-2-(1,2,3-thiadiazol-4-yl)acetate (anti isomer) (0.7 g.)was obtained as an oil.

I.R. ν_(max) ^(Film) : 1730, 1590 cm⁻¹.

N.M.R. δ_(ppm) (CDCl₃): 9.38 (1H, s), 4.47 (2H, q, J=7 Hz), 4.20 (3H,s), 1.40 (3H, t, J=7 Hz).

(3) 1N Aqueous solution of sodium hydroxide (6.7 ml.) was added to asolution of ethyl 2-methoxyimino-2-(1,2,3-thiadiazol-4-yl)acetate (synisomer) (1.2 g.) in methanol (10 ml.) and the mixture was stirred for1.5 hours at ambient temperature. Methanol was distilled off from thereaction mixture and water was added to the residue. The mixture waswashed with ether, adjusted to pH 1 with 10% hydrochloric acid andextracted with ethyl acetate. The extract was washed with a saturatedaqueous solution of sodium chloride and dried over magnesium sulfate.The solvent was distilled off to give prisms of2-methoxyimino-2-(1,2,3-thiadiazol-4-yl)acetic acid (syn isomer) (0.7g.), m.p. 110° to 113° C.

I.R. ν_(max) ^(Nujol) : 2750-2150, 1730, 1595 cm⁻¹.

N.M.R. δ_(ppm) (d₆ -DMSO): 9.47 (1H, s), 4.01 (3H, s).

EXAMPLE B

(1) Pulverized potassium carbonate (160 g.) was added to a solution ofethyl 2-hydroxyiminoacetoacetate (a mixture of syn and anti isomers)(152 g.) in acetone (500 ml.). Dimethyl sulfate (130 g.) was dropwiseadded thereto with stirring over 1 hour at 45° to 50° C. and the mixturewas stirred for 2 hours. An insoluble material was filtered off and thefiltrate was concentrated under reduced pressure. The filtered insolublematerial was dissolved in water (500 ml.) and this solution was added tothe residue. The mixture was extracted twice with ethyl acetate (300ml.). The extract was washed twice with water (200 ml.) and with asaturated sodium chloride aqueous solution (200 ml.) and dried overmagnesium sulfate. The solvent was distilled off under reduced pressureand the residue was distilled under reduced pressure to give colorlessoil of ethyl 2-methoxyiminoacetoacetate (a mixture of syn and antiisomers) (145.3 g.), bp 55° to 64° C./0.5 mm Hg.

I.R. ν_(max) ^(Film) : 1745, 1695, 1600 cm⁻¹.

N.M.R. δ_(ppm) (CDCl₃): 4.33 (4H, q, J=8 Hz), 4.08 (3H, s), 3.95 (3H,s), 2.40 (3H, s), 1.63 (3H, s), 1.33 (6H, t, J=8 Hz).

(2) Sulfuryl chloride (235 ml.) was dropwise added over 20 minutes withstirring and ice-cooling to a solution of ethyl2-methoxyiminoacetoacetate (syn isomer) (500 g.) in acetic acid (500ml.), and the mixture was stirred overnight under cooling with water.Nitrogen gas was introduced to the reaction mixture for 2 hours, and theresulting mixture was poured into water (2.5 l.) After extracting withmethylene chloride (500 ml.) and twice with methylene chloride (200ml.), the extracts were combined. The combined extract were washed witha saturated aqueous solution of sodium chloride, and adjusted to pH 6.5by adding water (800 ml.) and sodium bicarbonate. Methylene chloridelayer was separated, washed with an aqueous solution of sodium chlorideand dried over magnesium sulfate. The solvent was distilled off to giveethyl 2-methoxyimino-4-chloroacetoacetate (syn isomer) (559 g.).

I.R. ν_(max) ^(Film) : 1735, 1705 cm⁻¹.

(3) Ethyl 2-methoxyimino-4-chloroacetoacetate (syn isomer) (50 g.) wasadded over 3 minutes with stirring at ambient temperature to a solutionof thiourea (18.4 g.) and sodium acetate (19.8 g.) in a mixture ofmethanol (250 ml.) and water (250 ml.). After stirring for 35 minutes at40° to 45° C., the reaction mixture was cooled with ice and adjusted topH 6.3 with a saturated aqueous solution of sodium bicarbonate. Afterstirring for 30 minutes at the same temperature, precipitates werecollected by filtration, washed with water (200 ml.) and then withdiisopropyl ether (100 ml.), and dried to give colorless crystals ofethyl 2-methoxyimino-2-(2-amino-1,3-thiazol-4-yl)acetate (syn isomer)(37.8 g.), m.p. 161° to 162° C.

I.R. ν_(max) ^(Nujol) : 3400, 3300, 3150, 1725, 1630, 1559 cm⁻¹.

N.M.R. δ_(ppm) (CDCl₃): 6.72 (1H, s), 5.91 (2H, broad s), 4.38 (2H, q,J=7 Hz), 4.03 (3H, s), 1.38 (3H, t, J=7 Hz).

(4) Ethanol (10 ml.) was added to a suspension of ethyl2-methoxyimino-2-(2-amino-1,3-thiazol-4-yl)acetate (syn isomer) (2.2 g.)in a 1N aqueous solution of sodium hydroxide (12 ml.) and the mixturewas stirred for 15 hours at ambient temperature. The reaction mixturewas adjusted to pH 7.0 with 10% hydrochloric acid and ethanol wasdistilled off under reduced pressure. The residual aqueous solution waswashed with ethyl acetate, adjusted to pH 2.8 with 10% hydrochloric acidand stirred under ice-cooling to precipitate crystals. The crystals werecollected by filtration, washed with acetone and recrystallized fromethanol to give colorless needles of2-methoxyimino-2-(2-amino-1,3-thiazol-4-yl)acetic acid (syn isomer) (1.1g.)

I.R. ν_(max) ^(Nujol) : 3150, 1670, 1610, 1585 cm⁻¹.

N.M.R. δ_(ppm) (d₆ -DMSO): 7.20 (2H, broad s), 6.85 (1H, s), 3.83 (3H,s).

EXAMPLE C

(1) Sulfuryl chloride (35.2 g.) was added all at once to the stirredsolution of ethyl 2-ethoxyimino-3-oxobutyrate (syn isomer, 48.9 g.) inacetic acid (49 ml.) at room temperature, and stirred at the sametemperature for an hour. After adding the resultant solution into water(200 ml.), the solution was extracted with methylene chloride. Theextract was washed with a saturated aqueous solution of sodium chloride,neutralized with an aqueous solution of sodium bicarbonate and washedwith water. The solution was dried over magnesium sulfate andconcentrated under reduced pressure to give ethyl2-ethoxyimino-3-oxo-4-chlorobutyrate (syn isomer, 53.8 g.), pale yellowoil.

(2) A mixture of ethyl 2-ethoxyimino-3-oxo-4-chlorobutyrate (syn isomer38.7 g.), thiourea (13.2 g.), sodium acetate (14.3 g.), methanol (95ml.) and water (95 ml.) was stirred at 48° C. for 40 minutes. After theresultant solution was adjusted to pH 6.5 with an aqueous solution ofsodium bicarbonate, the appeared precipitates were collected byfiltration and washed with diisopropyl ether to give ethyl2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetate (syn isomer, 14.7 g.), mp130° to 131° C.

I.R. ν_(max) ^(Nujol) : 3450, 3275, 3125, 1715, 1620 cm⁻¹.

(3) Ethyl 2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetate (syn isomer, 5g.) was added to a mixture of 1N sodium hydroxide (45.9 ml.) and ethanol(30 ml.) and stirred at room temperature for 5 hours. After removingethanol from the resultant solution under reduced pressure, the residuewas dissolved in water (60 ml.) and adjusted to pH 2.0 with 10%hydrochloric acid. The solution was subjected to salting-out, and theprecipitates were collected by filtration and dried to give2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetic acid (syn isomer, 2.9 g.).

I.R. ν_(max) ^(Nujol) : 3625, 3225 (shoulder), 3100, 1650, 1615 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 1.20 (3H, t, J=7 Hz), 4.09 (2H, q, J=7 Hz), 6.82(1H, s), 7.24 (2H, broad s).

(4) 2-(2-Aminothiazol-4-yl)-2-ethoxyiminoacetic acid (syn isomer, 100g.), formic acid (85.5 g.) and acetic anhydride (190.1 g.) were treatedin a similar manner to that of Example F-(5) to give2-(2-formamidothiazol-4-yl)-2-ethoxyiminoacetic acid (syn isomer, 99.1g.).

I.R. ν_(max) ^(Nujol) : 3200, 3140, 3050, 1700 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.18 (3H, t, J=6 Hz), 4.22 (2H, q, J=6 Hz), 7.56(1H, s), 8.56 (1H, s), 12.62 (1H, broad s).

EXAMPLE D

(1) To a suspension of ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer,15 g.) and potassium carbonate (19.8 g.) in acetone (75 ml.) was addeddropwise propyliodide (16.2 g.) with stirring, and the mixture wasstirred at ambient temperature for 1,5 hours. The insoluble substancewas collected by filtration and washed with acetone. The washings andthe filtrate were combined and evaporated to dryness under reducedpressure. To the resultant residue was added water and the aqueoussolution was extracted twice with chloroform. The extract was washedwith an aqueous solution of sodium chloride, dried over magnesiumsulfate, and then evaporated to dryness under reduced pressure to giveethyl 3-oxo-2-propoxyiminobutyrate (syn isomer, 15.4 g.), oil.

(2) Ethyl 3-oxo-2-propoxyiminobutyrate (syn isomer, 15.4 g.) andsulfuryl chloride (10.6 g.) were dissolved in acetic acid (15.4 ml.),warmed at 35° to 40° C. for 10 minutes with stirring and then stirred atambient temperature for additional 6 hours. The reaction mixture waspoured into ice-water (200 ml.) and the resultant mixture was extractedtwice with chloroform. The extract was washed with an aqueous solutionof sodium chloride, twice a saturated aqueous solution of sodiumbicarbonate and once with water in turn, dried over magnesium sulfate,and then evaporated to dryness under reduced pressure to give ethyl4-chloro-3-oxo-2-propoxyiminobutyrate (syn isomer, 15.4 g.), oil.

I.R. ν_(max) ^(film) : 1740, 1710, 1695, 1455 cm⁻¹.

(3) Ethyl 4-chloro-3-oxo-2-propoxyiminobutyrate (syn isomer, 15.4 g.),thiourea (4.97 g.) and sodium acetate hydrate (8.89 g.) were dissolvedin a mixture of water (40 ml.) and ethanol (50 ml.), and stirred at 40°C. for an hour.

The reaction mixture was adjusted to pH 6.5 with a saturated aqueoussolution of potassium carbonate under cooling and stirred at the sametemperature for half an hour. The precipitating crystals were collectedby filtration, washed with water and diisopropyl ether, and then driedto give crystalline ethyl 2-(2-amino-4-thiazolyl)-2-propoxyiminoacetate(syn isomer, 10.55 g.), mp 142°-144° C.

I.R. ν_(max) ^(Nujol) : 3460, 3260, 3120, 1720, 1620, 1540 cm⁻¹.

N.M.R. δppm (d₆ -DMSO): 0.88 (3H, t, J=7 Hz), 1.27 (3H, t, J=6 Hz), 1.60(2H, sextet, J=7 Hz), 4.04 (2H, t, J=7 Hz), 4.28 (2H, q, J=6 Hz), 6.86(1H, s), 7.23 (2H, s).

(4) A solution of ethyl 2-(2-amino-4-thiazolyl)-2-propoxyiminoacetate(syn isomer, 10 g.) in a mixture of tetrahydrofuran (39 ml.), methanol(39ml.) and 1N sodium hydroxide (75.8 ml.) was stirred at 35° to 40° C.for 5 hours.

After the resultant solution was concentrated under reduced pressure,the aqueous residue was adjusted to pH 2.5 with 10% hydrochloric acid.The precipitates were collected by filtration and dried to give2-(2-amino-4-thiazolyl)-2-propoxyiminoacetic acid (synisomer, 6.2 g.),mp 161° C. (dec.)

I.R. ν_(max) ^(Nujol:) 3380, 3120 (broad), 1630, 1610, 1460 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 0.89 (3H, t, J=7 Hz), 1.63 (2H, sextet, J=7 Hz),4.05 (2H, t, J=7 Hz), 6.83 (1H, s), 6.9-8.8 (3H, broad).

(5) 2-(2-Aminothiazol-4-yl)-2-n-propoxyiminoacetic acid (syn isomer,21.8 g.), acetic anhydride (38.8 g.) and formic acid (17.5 g.) weretreated in a similar manner to that of Example F-(5), and then theobtained oil was triturated with diisopropyl ether to give2-(2-formamidothiazol-4-yl)-2-n-propoxyiminoacetic acid (syn isomer,19.2 g.), mp. 164° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3200, 3120, 3050, 1700, 1550 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.92 (3H, t, J=7 Hz), 1.67 (2H, sextet, J=7 Hz),4.12 (2H, t, J=7 Hz), 7.53 (1H, s), 8.54 (1H, s).

EXAMPLE E

(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 30 g.), iso-propyliodide (32.5 g.), potassium carbonate (39.5 g.) and acetone (150 ml.)were treated in a similar manner to that of Example D-(1) to give ethyl2-iso-propoxyimino-3-oxobutyrate (syn isomer, 35.4 g.), oil.

I.R. ν_(max) ^(Film) : 1745, 1690, 1600 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 1.33 (3H, t, J=7 Hz), 1.35 (6H, d, J=6 Hz), 2.32(3H, s), 4.1˜4.7 (3H, m).

(2) Ethyl 2-iso-propoxyimino-3-oxobutyrate (syn isomer 35.4 g.),sulfuryl chloride (24.5 g.) and acetic acid (35.4 ml.) were treated in asimilar manner to that of Example D-(2) to give ethyl4-chloro-3-oxo-2-iso-propoxyiminobutyrate (syn isomer, 41.5 g.), oil.

I.R. ν_(max) ^(Film) : 1745, 1715, 1375 cm⁻¹.

(3) Ethyl 4-chloro-3-oxo-2-iso-propoxyiminobutyrate (syn isomer, 41.5g.), thiourea (13.4 g.), sodium acetate (14.4 g.), water (110 ml.) andethanol (110 ml.) were treated in a similar manner to that of ExampleD-(3) to give ethyl 2-(2-aminothiazol-4-yl)-2-iso-propoxyiminoacetate(syn isomer, 27.3 g.), mp. 162° to 164° C.

I.R. ν_(max) ^(Nujol) : 3460, 3430, 3260, 3150, 1725, 1615 1540 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.17 (6H, d, J=6 Hz), 1.24 (3H, t, J=7 Hz),4˜4.7 (3H, m), 6.86 (1H, s), 7.24 (2H, s).

(4) Ethyl 2-(2-aminothiazol-4-yl)-2-iso-propoxyiminoacetate (syn isomer,26.8 g.), 1N aqueous solution of sodium hydroxide (156 ml.), methanol(156 ml.) and tetrahydrofuran (100 ml.) were treated in a similar mannerto that of Example D-(4) to give2-(2-aminothiazol-4-yl)-2-iso-propoxyiminoacetic acid (syn isomer, 15.3g.), mp. 151° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3610, 3580, 3080, 1650, 1610 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.22 (6H, d, J=6 Hz), 4.33 (1H, quintet, J=6Hz), 6.80 (1H, s), 7.22 (2H, broad s).

(5) 2-(2-Aminothiazol-4-yl)-2-iso-propoxyiminoacetic acid (syn isomer, 4g.), acetic anhyddride (7.6 g.) and formic acid (3.4 g.) were treated ina similar manner to that of Example F-(5) to give2-(2-formamidothiazol-4-yl)-2-iso-propoxyiminoacetic acid (syn isomer,3.75 g.), mp. 168° to 169° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3200, 3130, 1710, 1600, 1560 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.26 (6H, d), 4.4 (1H, m), 7.54 (1H, s), 8.52(1H, s), 12.56 (1H, broad s).

EXAMPLE F

(1) n-Butyl iodide (46.9 g.) was added dropwise to a stirred suspensionof ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 40 g.), potassiumcarbonate (52.7 g.) and acetone (200 ml.) under ice-cooling over 5minutes, and stirred at room temperature for 4 hours. The resultantsolution was filtered, and washed with acetone. The filtrate and washingsolution were combined together and concentrated in vacuo. After addingwater (300 ml.) to the residue, the solution was extracted withmethylene chloride three times. The solution was washed with a saturatedaqueous solution of sodium chloride, dried over magnesium sulfate andconcentrated in vacuo to give ethyl 2-n-butoxyimino-3-oxobutyrate (synisomer, 48.8 g.), oil.

I.R. ν_(max) ^(Film) : 1750, 1700, 1470, 1370, 1320 cm⁻¹.

(2) A solution of ethyl 2-n-butoxyimino-3-oxobutyrate (syn isomer, 48.8g.), sulfuryl chloride (31.5 g.) and acetic acid (48.8 ml.) was stirredat 40° C. for 10 minutes and further at room temperature for 5.5 hours.After water (300 ml.) was added to the resultant solution under icecooling, the solution was extracted with methylene chloride three times.The extract was washed with water, an aqueous solution of sodiumbicarbonate and a saturated aqueous solution of sodium chloride in turn,and dried over magnesium sulfate. The solution was concentrated in vacuoto give ethyl 2-n-butoxyimino-4-chloro-3-oxobutyrate (syn isomer, 52.1g.), oil.

I.R. ν_(max) ^(Film) : 1740, 1710, 1470, 1370 cm⁻¹.

(3) A solution of ethyl 2-n-butoxyimino-4-chloro-3-oxobutyrate (synisomer, 52.1 g.), thiourea (15.9 g.), sodium acetate 3 hydrate (28.4g.), water (130 ml.) and ethanol (180 ml.) was stirred at 40° C. for1.25 hours. The resultant solution was adjusted to pH 6.5 with anaqueous solution of sodium carbonate under ice cooling, and stirred for20 minutes under ice cooling. The precipitates were collected byfiltration, and washed with water and diisopropyl ether in turn to giveethyl 2-(2-aminothiazol-4-yl)-2-n-butoxyiminoacetate (syn isomer, 36.1g.), mp 126° to 128° C.

I.R. ν_(max) ^(Nujol) : 3460, 3370, 3230, 1720, 1620, 1550 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6-2.0 (6H, m), 1.28 (3H, t, J=7 Hz), 4.12 (3H,t, J=6 Hz), 4.31 (2H, q, J=7 Hz), 6.89 (1H, s), 7.24 (2H, s).

(4) A solution of ethyl 2-(2-aminothiazol-4-yl)-2-n-butoxyiminoacetate(syn isomer, 36 g.), methanol (133 ml.), tetrahydrofuran (133 ml.) and2N aqueous solution of sodium hydroxide (133 ml.) was stirred at 30° C.for 5 hours. After the resultant solution was concentrated in vacuo, theresidue was dissolved in water. The solution was adjusted to pH 7 with10% hydrochloric acid and treated with activated charcoal. The solutionwas adjusted to pH 2.0 with 10% hydrochloric acid and stirred for 20minutes under ice cooling. The precipitates were collected byfiltration, washed with water and acetone in turn, and dried to give2-(2-aminothiazol-4-yl)-2-n-butoxyiminoacetic acid (syn isomer, 25.4g.).

I.R. ν_(max) ^(Nujol) : 3325, 3190, 1660, 1620 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.88 (3H, t, J=7 Hz), 1.0-1.9 (4H, m), 4.06 (2H,t, J=7 Hz), 6.81 (1H, s), 7.21 (2H, broad s).

(5) Formic acid (18.95 g.) was added dropwise to acetic anhydride (42.0g.) under stirring at room temperature over 5 minutes, and stirred at50° C. for an hour. 2-(2-Aminothiazol-4-yl)-2-n-butoxyiminoacetic acid(syn isomer, 25 g.) was added to the solution under ice cooling, andstirred at room temperature for 3 hours and additionally at 30° C. foran hour. After concentrating the resultant solution in vacuo, theresidue was dissolved in diethyl ether. The solution was washed withwater and a saturated aqueous solution of sodium chloride in turn, driedover magnesium sulfate and concentrated in vacuo. The obtained oil wastriturated with a solution of n-hexane (1 part) and diisopropyl ether (1part), and collected by filtration to give2-(2-formamidothiazol-4-yl)-2-n-butoxyiminoacetic acid (syn isomer, 20.1g.).

I.R. ν_(max) ^(Nujol) : 3350, 3160, 3050, 1700, 1680, 1570 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.91 (3H, t, J=6 Hz), 1.0-2.2 (4H, m), 4.18 (2H,t, J=6 Hz), 7.57 (1H, s), 8.59 (1H, s), 12.66 (1H, broad s).

EXAMPLE G

(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 40 g.),N,N-dimethylformamide (200 ml.), potassium carbonate (52.7 g.) andiso-butyl bromide (34.94 g.) were treated in a similar manner to that ofExample F-(1) to give ethyl 2-iso-butoxyimino-3-oxo-butyrate (synisomer, 42 g.).

I.R. ν_(max) ^(Nujol) : 1740, 1670 (broad) cm⁻¹.

(2) Ethyl 2-iso-butoxyimino-3-oxobutyrate (syn isomer, 42 g.), aceticacid (42 ml.) and sulfuryl chloride (27.1 g.) were treated in a similarmanner to that of Example F-(2) to give ethyl2-iso-butoxyimino-4-chloro-3-oxobutyrate (syn isomer, 31.9 g.).

I.R. ν_(max) ^(Nujol) : 1750, 1720, 1680 cm⁻¹.

(3) Ethyl 2-iso-butoxyimino-4-chloro-3-oxobutyrate (syn isomer, 31.9g.), thiourea (9.72 g.), sodium acetate 3-hydrate (17.4 g.), ethanol(120 ml.) and water (80 ml.) were treated in a similar manner to that ofExample F-(3) to give ethyl2-(2-aminothiazol-4-yl)-2-iso-butoxyiminoacetate (syn isomer, 17.6 g.),mp 122° to 124° C.

I.R. ν_(max) ^(Nujol) : 3470, 3260, 3120, 1730, 1620, 1545 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.86 (6H, d, J=7 Hz), 1.28 (3H, t, J=7 Hz),1.6-2.2 (1H, m), 3.86 (2H, d, J=7 Hz), 4.28 (2H, q, J=7 Hz), 6.86 (1H,s), 7.22 (2H, s).

(4) Ethyl 2-(2-aminothiazol-4-yl)-2-iso-butoxyiminoacetate (syn isomer,19.6 g.), 2N aqueous solution of sodium hydroxide (72.2 ml.), methanol(72.2 ml.) and tetrahydrofuran (72.2 ml.) were treated in a similarmanner to that of Example F-(4) to give2-(2-aminothiazol-4-yl)-2-isobutoxyiminoacetic acid (syn isomer, 16.1g.), mp 180° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3375, 3300, 3130, 3050, 1640 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.91 (6H, d, J=7 Hz), 1.5-2.3 (1H, m), 3.90 (2H,d, J=7 Hz), 6.87 (1H, s), 7.26 (2H, broad s).

(5) 2-(2-Aminothiazol-4-yl)-2-iso-butoxyiminoacetic acid (syn isomer,11.5 g.) acetic anhydride (19.3 g.) and formic acid (8.7 g.) weretreated in a similar manner to that of Example F-(5) to give2-(2-formamidothiazol-4-yl)-2-iso-butoxyiminoacetic acid (syn isomer,11.15 g.), mp 163° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3175, 3110, 3050, 1695, 1550 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.91 (6H, d, J=7 Hz), 1.7-2.3 (1H, m), 3.92 (2H,d, J=7 Hz), 7.52 (1H, s), 8.52 (1H, s), 12.58 (1H, broad s).

EXAMPLE H

(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 30 g.),N,N-dimethylformamide (100 ml.), potassium carbonate (39.5 g.) andcyclohexyl bromide (31.1 g.) were treated in a similar manner to that ofExample F-(1) to give ethyl 2-cyclohexyloxyimino-3-oxobutyrate (synisomer, 41.8 g.), oil.

I.R. ν_(max) ^(Film) : 1740, 1680 cm⁻¹.

(2) Ethyl 2-cyclohexyloxyimino-3-oxobutyrate (syn isomer, 41.3 g.),acetic acid (41.3 ml.) and sulfuryl chloride (23.8 g.) were treated in asimilar manner to that of Example F-(2) to give ethyl4-chloro-2-cyclohexyloxyimino-3-oxobutyrate (syn isomer, 27.8 g.), oil.

I.R. ν_(max) ^(Film) : 1745, 1715, 1680 cm⁻¹.

(3) Ethyl 4-chloro-2-cyclohexyloxyimino-3-oxobutyrate (syn isomer, 27.8g.), thiourea (7.7 g.), sodium acetate 3-hydrate (13.7 g.), water (70ml.) and ethanol (140 ml.) were treated in a similar manner to that ofExample F-(3) to give ethyl2-(2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetate (syn isomer, 3.6g.), mp. 125° to 126° C.

I.R. ν_(max) ^(Nujol) : 3430, 3250, 3160, 3130, 1715, 1635 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.28 (3H, t, J=7 Hz), 1.0˜2.2 (10H, m), 4.22(1H, m), 4.32 (2H, q, J=7 Hz), 6.88 (1H, s), 7.24 (2H, broad s).

(4) Ethyl 2-(2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetate (synisomer, 3.5 g.), 2N aqueous solution of sodium hydroxide (11.8 ml.),methanol (11.8 ml.) and tetrahydrofuran (11.8 ml.) were treated in asimilar manner to that of Example F-(4) to give2-(2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetic acid (syn isomer, 2.1g.), mp. 148° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3110, 1630, 1450 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.8˜2.3 (10H, m), 4.14 (1H, m), 6.86 (1H, s),7.5 (2H, broad s).

(5) 2-(2-Aminothiazol-4-yl)-2-cyclohexyloxyiminoacetic acid (syn isomer,1.5 g.), acetic anhydride (2.27 g.) and formic acid (1.03 g.) weretreated in a similar manner to that of Example F-(5), and the oilobtained was suspended in an aqueous solution of sodium bicarbonate. Thesuspension was adjusted to pH 3.5 with 10% hydrochloric acid. Theprecipitates were collected by filtration, washed with water and driedto give 2-(2-formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetic acid (synisomer, 1.0 g.), mp. above 230° C.

I.R. ν_(max) ^(Nujol) : 3175, 3100, 3060, 1680 cm⁻¹.

EXAMPLE I

(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 56.7 g.),N,N-dimethylformamide (280 ml.), potassium carbonate (72.3 g.) andpropargyl bromide (43 g.) were treated in a similar manner to that ofExample F (1) to give ethyl 2-propargyloxyimino-3-oxobutyrate (synisomer, 71.2 g.).

I.R. ν_(max) ^(Film) : 3280, 3220, 2120, 1735, 1670 cm⁻¹.

(2) Ethyl 2-propargyloxyimino-3-oxobutyrate (syn isomer, 71.2 g.),acetic acid (81 ml.) and sulfuryl chloride (50.2 g.) were treated in asimilar manner to that of Example F-(2) to give ethyl4-chloro-3-oxo-2-propargyloxyiminobutyrate (syn isomer, 61.6 g.), oil.

I.R. ν_(max) ^(Film) : 3300, 2130, 1745, 1720, 1675 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 1.39 (3H, t, J=7 Hz), 2.57 (1H, t, J=2 Hz), 4.36(2H, q, J=7 Hz), 4.56 (2H, s), 4.86 (2H, d, J=2 Hz).

(3) Ethyl 4-chloro-3-oxo-2-propargyloxyiminobutyrate (syn isomer, 61g.), thiourea (20 g.), sodium acetate 3-hydrate (35.8 g.), water (150ml.) and ethanol (180 ml.) were treated in a similar manner to that ofExample F-(3) to give ethyl2-(2-aminothiazol-4-yl)-2-proparglyoxyiminoacetate (syn isomer, 35.6g.).

I.R. ν_(max) ^(Nujol) : 3290, 2220, 1729 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.28 (3H, t, J=7 Hz), 3.49 (1H, t, J=3 Hz), 4.31(2H, q, J=7 Hz), 4.76 (2H, d, J=3 Hz), 6.95 (1H, s), 7.29 (2H, s).

(4) Ethyl 2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetate (synisomer, 2.8 g.), methanol (23 ml.), tetrahydrofuran (20 ml.) and 1Naqueous solution of sodium hydroxide (22.17 ml.) were treated in asimilar manner to that of Example F-(4) to give2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetic acid (syn isomer,1.924 g.).

I.R. ν_(max) ^(Nujol) : 2190, 1740.

N.M.R. δ(DMSO-d₆, ppm): 3.47 (1H, t, J=1.5 Hz), 4.74 (2H, d, J=1.5 Hz),6.90 (1H, s).

EXAMPLE J

(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 40 g.),N,N-dimethylformaide (200 ml.), potassium carbonate (52 g.) and n-hexylbromide (41.4 g.) were treated in a similar manner to that of ExampleF-(1) to give ethyl 2-n-hexyloxyimino-3-oxobutyrate (syn isomer, 60.7g.). oil.

I.R. ν_(max) ^(Film) : 1740, 1705, 1700 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 0.6˜2.1 (14H, m), 2.37 (3H, s), 4.1˜4.6 (4H, m).

(2) Ethyl 2-n-hexyloxyimino-3-oxobutyrate (syn isomer, 60.7 g.), aceticacid (61 ml.) and sulfuryl chloride (34.7 g.) were treated in a similarmanner to that of Example F-(2) to give ethyl2-n-hexyloxyimino-4-chloro-3-oxobutyrate (syn isomer, 55.6 g.).

I.R. ν_(max) ^(Film) : 1740, 1720, 1470 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 0.6˜2.2 (14H, m), 4.1˜4.6 (4H, m), 4.47 (2H, s).

(3) Ethyl 2-n-hexyloxyimino-4-chloro-3-oxobutyrate (syn isomer, 55.6g.), thiourea (15.2 g.), sodium acetate 3-hydrate (27.2 g.), ethanol(280 ml.) and water (140 ml.) were treated in a similar manner to thatof Example F-(3) to give ethyl2-(2-aminothiazol-4-yl)-2-n-hexyloxyiminoacetate (syn isomer, 29.3 g.),mp 77° to 78° C.

I.R. ν_(max) ^(Nujol) : 3460, 3250, 3140, 1720, 1535 cm⁻¹.

N.M.R. δ(DMSO-d₆): 0.85 (3H, t, J=6 Hz), 1,0˜1.9 (11H, m), 2.07 (2H, t,J=6 Hz), 2,26 (2H, q, J=7 Hz), 6.85 (1H, s), 7.22 (2H, s).

(4) Ethyl 2-(2-aminothiazol-4-yl)2-n-hexyloxyiminoacetate (syn isomer,29.1 g.), methanol (97.2 ml.), 2N aqueous solution of sodium hydroxide(97.2 ml.) and tetrahydrofuran (50 ml.) were treated in a similar mannerto that of Example F-(4) to give2-(2-aminothiazol-4-yl)-2-n-hexyloxyiminoacetic acid (syn isomer, 24.0g.), mp. 174° C. (dec.).

I.R. ν_(max) ^(Nujol) : 1660, 1625, 1425 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm) 0.6˜2.1 (11H, m), 4.07 (2H, t, J=6 Hz), 6.83 (1H,s), 7.19 (2H, s).

EXAMPLE K

(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 40 g.),N,N-dimethylformamide (200 ml.), potassium carbonate (52 g.) and pentylbromide (37.9 g.) were treated in a similar manner to that of ExampleF-(1) to give ethyl 2-pentyloxyimino-3-oxobutyrate (syn isomer, 57.5g.), oil.

I.R. ν_(max) ^(Film) : 1745, 1680, 1470 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 0.7-2.2 (12H, m), 2.36 (3H, s), 4.1-4.6 (4H, m).

(2) Ethyl 2-pentyloxyimino-3-oxobutyrate (syn isomer, 57.5 g.), aceticacid (58.5 ml.) and sulfuryl chloride (20.9 ml.) were treated in asimilar manner to that of Example F-(2) to give ethyl2-pentyloxyimino-4-chloro-3-oxobutyrate (syn isomer, 51.1 g.), oil.

I.R. ν_(max) ^(Film) : 1750, 1715, 1470 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 0.7-2.1 (11H, m), 4.1-4.6 (4H, m), 4.48 (2H, s).

(3) Ethyl 2-pentyloxyimino-4-chloro-3-oxobutyrate (syn isomer, 51.1 g.),thiourea (14.7 g.), sodium acetate trihydrate (26.4 g.), ethanol (175ml.) and water (125 ml.) were treated in a similar manner to that ofExample F-(3) to give ethyl2-(2-aminothiazol-4-yl)-2-pentyloxyiminoacetate (syn isomer, 28.7 g.),mp 86° to 88° C.

I.R. ν_(max) ^(Nujol) : 3450, 3250, 3130, 1715, 1535 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6-2.0 (12H, m), 4.11 (2H, t, J=6 Hz), 4.32(2H, q, J=7 Hz), 6.90 (1H, s), 7.25 (2H, s).

(4) Ethyl 2-(2-aminothiazol-4-yl)-2-pentyloxyiminoacetate (syn isomer,28.6 g.), 2N aqueous solution of sodium hydroxide (100.2 ml.), methanol(100 ml.) and tetrahydrofuran (100 ml.) were treated in a similar mannerto that of Example F-(4) to give2-(2-aminothiazol-4-yl)-2-pentyloxyiminoacetic acid (syn isomer, 22.4g.), mp 176° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3160, 1655, 1620, 1460 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6-2.2 (9H, m), 4.07 (2H, t, J=6 Hz), 6.82 (1H,s), 7.20 (2H, s).

(5) 2-(2-Aminothiazol-4-yl)-2-pentyloxyiminoacetic acid (syn isomer, 15g.), acetic anhydride (23.8 g.) and formic acid (10.7 g.) were treatedin a similar manner to that of Example F-(5) to give2-(2-formamidothiazol-4-yl)-2-pentyloxyiminoacetic acid (syn isomer,14.7 g.), mp 125° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3200, 3140, 1700, 1565 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6-2.0 (9H, m), 4.13 (2H, t, J=6 Hz), 7.53 (1H,s), 7.54 (1H, s), 12.66 (1H, s).

EXAMPLE L

(1) Allyl bromide (2.91 g.) was added dropwise to a stirred suspensionof ethyl 2-(2-tritylaminothiazol-4-yl)-2-hyroxyiminoacetate (syn isomer,10 g.), N,N-dimethylformamide (100 ml.) and potassium carbonate (4.54g.) under ice cooling over 5 minutes, and stirred at the sametemperature for 4 hours. After adding water (200 ml.) to the resultantsolution, the solution was extracted with diethyl ether twice. Theextract was washed with a saturated aqueous solution of sodium chlorideand dried over magnesium sulfate. The solution was concentrated invacuo, and the residue was triturated with a solution of n-hexane anddiethyl ether. The precipitates were collected by filtration to giveethyl 2-(2-tritylaminothiazol-4-yl)-2-allyloxyiminoacetate (syn isomer,9.4 g.), mp. 130° to 132° C.

I.R. ν_(max) ^(Nujol) : 3380, 1735, 1520, 1500 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.08 (3H, t, J=7 Hz), 3.96 (2H, q, J=7 Hz), 4.54(2H, broad d, J=5 Hz), 5.0˜5.5 (2H, m), 5.6˜6.3 (1H, m), 6.90 (15H,broad s), 7.74 (1H, s).

(2) A solution of ethyl2-(2-tritylaminothiazol-4-yl)-2-allyloxyiminoacetate (syn isomer, 8.7g.), 50% formic acid (42.5 ml.) and tetrahydrofuran (42.5 ml.) wasstirred at 60° C. for 40 minutes. After concentrating the resultantsolution in vacuo, the residue was dissolved in ethyl acetate, washedwith an aqueous solution of sodium bicarbonate and a saturated aqueoussolution of sodium chloride in turn, and dried over magnesium sulfate.After concentrating the resultant solution in vacuo, the residue wassubjected to column chromatography on silica gel with benzene and ethylacetate in turn, to give ethyl2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetate (syn isomer, 3.7 g.), mp.102° to 104° C.

I.R. ν_(max) ^(Nujol) : 3460, 3260, 3130, 1725, 1620, 1540, 1460 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.25 (3H, t, J=7 Hz), 4.30 (2H, q, J=7 Hz), 4.61(2H, dd, J=5 Hz, 1 Hz), 5.0˜5.5 (2H, m), 5.6˜6.5 (1H, m), 6.95 (1H, s),7.28 (2H, s).

(3) A solution of ethyl 2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetate(syn isomer, 3.6 g.), 2N aqueous solution of sodium hydroxide (14.1ml.), tetrahydrofuran (14.1 ml.) and methanol (15 ml.) was stirred at40° C. for 1.5 hours. The resultant solution was concentrated in vacuo,and the residue was dissolved in water. After the solution was adjustedto pH 2.8 with 10% hydrochloric acid under ice cooling, the precipitateswere collected by filtration, washed with water and acetone in turn anddried to give 2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetic acid (synisomer, 1.91 g.), mp 187° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3350, 1630, 1580, 1460 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 4.61 (2H, d, J=6 Hz), 5.1˜5.5 (2H, m), 5.7˜6.2(1H, m), 6.84 (1H, s), 7.25 (2H, broad s).

EXAMPLE M

(1) Propargyl bromide (4.16 g.) was added to a suspension of ethyl2-(2-tritylaminothiazol-4-yl)-2-hydroxyiminoacetate (syn isomer, 10 g.),potassium carbonate (4.84 g.) and N,N-dimethylformamide (22 ml.) underatmosphere of nitrogen gas and stirred at room temperature for 100minutes. The insoluble substance was filtered off and washed with alittle of N,N-dimethylformamide. The filtrate and washing solution werecombined together, and water (400 ml.) was added to the solution. Afterthe suspension was extracted with ethyl acetate (400 ml.), the extractwas washed with a saturated aqueous solution of sodium chloride anddried over magnesium sulfate. After treating the solution with activatedcharcoal, the solution was concentrated in vacuo. The residue wastriturated with diisopropyl ether. The precipitates were collected byfiltration, and washed with diisopropyl ether to give ethyl2-(2-tritylaminothiazol-4-yl)-2-propargyloxyiminoacetate (syn isomer,8.34 g.).

I.R. ν_(max) ^(Nujol) : 3290, 2225, 1735 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.12 (3H, t, J=7 Hz), 3.47 (1H, t, J=3 Hz), 3.97(2H, q, J=7 Hz), 4.67 (2H, d, J=3 Hz), 6.95 (1H, s), 7.26 (15H, s), 8.77(1H, s).

(2) 50% Formic acid (41 ml.) was added to a solution of ethyl2-(2-tritylaminothiazol-4-yl)-2-propargyloxyiminoacetate (syn isomer,8.2 g.) and tetrahydrofuran (41 ml.), and stirred at 60° C. for an hour.The resultant solution was concentrated to a half of initial volumeunder reduced pressure, and the precipitates were collected byfiltration and washed with diisopropyl ether. The filtrate and washingsolution were combined together and concentrated in vacuo. The residuewas added to ethyl acetate (200 ml.) under stirring. The insolublesubstance was collected by filtration, and washed with diethyl ether togive ethyl 2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetate (synisomer, 0.3 g.). The filtrate and ethyl acetate washing solution werecombined together, washed with a saturated aqueous solution of sodiumbicarbonate and a saturated aqueous solution of sodium chloride twice inturn, and dried over magnesium sulfate. The solution was treated withactivated charcoal and concentrated in vacuo. The residue was dried invacuo after adding benzene. The residue was subjected to columnchromatography on silica gel with benzene and ethyl acetate in turn. Theeluate was concentrated in vacuo, and the residue was triturated withdiisopropyl ether. The precipitates were collected by filtration, washedwith diisopropyl ether to give the same compound as mentioned above (synisomer, 2.658 g.). The I.R. spectrum and N.M.R. spectrum are the same asthose of the compound obtained in Example I-(3).

EXAMPLE N

Sodium bicarbonate (0.84 g.) was added to a suspension of2-(2-formamidothiazol-4-yl)oxalic acid (2 g.) in water (120 ml.) toprepare a solution. Ethyl 2-aminoxyacetate hydrochloride (4.56 g.) wasadded to the solution and stirred at room temperature for 3 hours whileadjusting to pH 6 with sodium bicarbonate. The resultant solution wasadjusted to pH 1.5 with hydrochloric acid, salted out and extracted withethyl acetate three times. The extract was dried over magnesium sulfateand concentrated in vacuo. The residue was pulverized with diethylether, and the precipitates were collected by filtration and dried togive 2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetic acid(syn isomer, 1.44 g.), mp 112° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3150, 1740, 1670, 1550 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.23 (3H, t, J=7 Hz), 4.16 (2H, q, J=7 Hz), 4.77(2H, s), 7.56 (1H, s), 8.54 (1H, s).

EXAMPLE O

(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 60 g.),1-bromo-2-chloroethane (54.1 g.), potassium carbonate (78 g.) andN,N-dimethylformamide (200 ml.) were treated in a similar manner to thatof Example F-(1) to give ethyl 2-(2-chloroethoxyimino)-3-oxobutyrate(syn isomer, 83.6 g.), oil.

I.R. ν_(max) ^(Film) : 1740, 1680, 1430 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 1.34 (3H, t, J=7 Hz), 2.34 (3H, s), 3.72 (2H, t,J=6 Hz), 4.28 (2H, q, J=7 Hz), 4.46 (2H, t, J=6 Hz)

(2) Ethyl 2-(2-chloroethoxyimino)-3-oxobutyrate (syn isomer, 83.6 g.),sulfury chloride (52.4 g.) and acetic acid (83.6 ml.) were treated in asimilar manner to that of Example F-(2) to give Ethyl2-(2-chloroethoxyimino)-3-oxo-4-chlorobutyrate (syn isomer, 68 g.), oil.

I.R. ν_(max) ^(Film) : 1740, 1720 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 1.32 (3H, t, J=7 Hz), 3.70 (2H, t, J=6 Hz), 4.29(2H, q, J=7 Hz), 4.47 (2H, s), 4.48 (2H, t, J=6 Hz).

(3) Ethyl 2-(2-chloroethoxyimino)-3-oxo-4-chlorobutyrate (syn isomer, 68g.), thiourea (20.2 g.), sodium acetate trihydrate (36.2 g.), ethanol(270 ml.) and water (170 ml.) were treated in a similar manner to thatof Example F-(3) to give ethyl2-(2-aminothiazol-4-yl)-2-(2-chloroethoxyimino)acetate (syn isomer, 33.7g.), mp 126° to 128° C.

I.R. ν_(max) ^(Nujol) : 3440, 3260, 3140, 1725, 1620, 1540 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.30 (3H, t, J=7 Hz), 3.78 (2H, t, J=6 Hz),4.1-4.6 (4H, m), 6.96 (1H, s), 7.27 (2H, s).

(4) Ethyl 2-(2-aminothiazol-4-yl)-2-(2-chloroethoxyimino)acetate (synisomer, 30.5 g.), 1N aqueous solution of sodium hydroxide (220 ml.),methanol (110 ml.) and tetrahydrofuran (140 ml.) were treated in asimilar manner to that of Example F-(4) to give2-(2-aminothiazol-4-yl)-2-(2-chloroethoxyimino)acetic acid (syn isomer,23.4 g.), mp 201° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3210, 3100, 1640, 1620, 1580 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.83 (2H, t, J=6 Hz), 4.36 (2H, t, J=6 Hz), 6.92(1H, s), 7.30 (2H, s).

(5) 2-(2-Aminothiazol-4-yl)-2-(2-chloroethoxyimino)acetic acid (synisomer, 15 g.), acetic anhydride (24.5 g.), formic acid (11.0 g.) andtetrahydrofuran (50 ml.) were treated in a similar manner to that ofExample F-(5) to give2-(2-formamidothiazol-4-yl)-2-(2-chloroethoxyimino)acetic acid (synisomer, 13.4 g.), mp 155° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3100, 1740, 1690, 1660 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.87 (2H, t, J=6 Hz), 4.40 (2H, t, J=6 Hz), 7.60(1H, s), 8.56 (1H, s), 12.62 (1H, broad s).

EXAMPLE P

A suspension of 2-(2-formamidothiazol-4-yl)oxalic acid (3.0 g.) inmethanol (60 ml.) and water (60 ml.) was adjusted to pH 8 with 1Naqueous solution of sodium hydroxide under stirring.2,2,2-Trifluoroethoxyamine hydrochloride (2.24 g.) was added to thesolution, and the solution was adjusted to pH 2.5 to 3 with 1N aqueoussolution of sodium hydroxide. After the solution was stirred at roomtemperature for 1.5 hours, methanol was removed from the resultantsolution under reduced pressure. The concentrated aqueous solution wasadjusted to pH 7 with 1N aqueous solution of sodium hydroxide and washedwith ethyl acetate. Ethyl acetate was added to the aqueous solution andadjusted to pH 1.5 with 10% hydrochloric acid, and then extracted withethyl acetate. The aqueous layer was extracted again with ethyl acetate.The extracts were combined, washed with a saturated aqueous solution ofsodium chloride and dried over magnesium sulfate. The solution wasconcentrated in vacuo to give 2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetic acid (syn isomer, 2.4 g.), mp 162°to 163° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3200, 1700, 1600, 1560 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 4.83 (2H, q, J=8.5 Hz), 7.65 (1H, s), 8.58 (1H,s), 12.60 (1H, broad s).

EXAMPLE Q

2-(2-Formamidothiazol-4-yl)oxalic acid (10 g.), sodium bicarbonate (4.2g.) and tert-butyl 2-aminooxyacetate (8.1 g.) were treated in a similarmanner to that of Example N to give an oil. The oil was triturated withn-hexane and the precipitates were collected by filtration and dried togive 2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoaceticacid (syn isomer, 11.3 g.), mp 117° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3180, 3140, 1750, 1690, 1630 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.46 (9H, s), 4.66 (2H, s), 7.56 (1H, s), 8.56(1H, s), 12.67 (1H, broad s).

EXAMPLE 1

(1) N,N-Dimethylformamide (0.16 g.) and phosphorous oxychloride (0.34g.) were mixed to prepare Vilsmeier reagent in a usual manner, and theresultant Vilsmeier reagent was suspended in dry ethyl acetate. To thesuspension was added 2-(2-formamido-4-thiazolyl)-2-methoxyiminoaceticacid (syn isomer, 0.46 g.) under ice-cooling with stirring, and then thesolution was stirred at the same temperature for 30 minutes to preparethe activated acid solution. p-Nitrobenzyl7-amino-3-chloro-3-cephem-4-carboxylate hydrochloride (0.81 g.) wasdissolved in a solution of trimethylsilylacetamide (2.10 g.) in ethylacetate (200 ml.). To the solution was added the activated acid solutionobtained above all at once at -20° C., and the solution was stirred at-20°˜-5° C. for 1.5 hours. After water and ethyl acetate (100 ml.) wereadded to the resultant solution at -20° C. the insoluble product wasseparated by filtration, washed with water and acetone in turn and thendried to give p-nitrobenzyl7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylate(syn isomer, 0.6 g.).

After ethyl acetate was removed from the above filtrate, the aqueouslayer was extracted with ethyl acetate (50 ml.) twice. The ethyl acetatelayer and the extract were combined together, washed with 10%hydrochloric acid, a saturated aqueous solution of sodium bicarbonateand a saturated aqueous solution of sodium chloride in turn and thendried over magnesium sulfate. After removing ethyl acetate from thesolution, diethyl ether was added to the residue. The insoluble productwas collected by filtration to give the same object compound (0.25 g.),m.p. 226° to 228° C. (dec.), Total yield 0.85 g.

I.R. ν_(max) ^(Nujol) : 3250, 1780, 1720, 1685, 1645, 1605, 1550, 1520cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.45 (2H, broad s), 3.93 (3H, s), 5.35 (1H, d,J=5 Hz), 5.50 (2H, s), 5.95 (1H, dd, J=5,8 Hz), 7.43 (1H, s), 7.72 (2H,d, J=9 Hz), 8.28 (2H, d, J=9 Hz), 8.55 (1H, s), 9.80 (1H, d, J=8 Hz).

(2) p-Nitrobenzyl7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylate(syn isomer, 0.8 g.) was dissolved in a mixed solution of methanol (30ml.) and tetrahydrofuran (60 ml.). After adding 10% palladium carbon(0.4 g.) to the solution, the mixture was subjected to catalyticreduction at room temperature under atmospheric pressure. The catalystwas filtered off, and the filtrate was concentrated under reducedpressure. Water (30 ml.) was added to the residue and the mixture wasadjusted to pH 7.5 with an aqueous solution of sodium bicarbonate. Afterremoving the insoluble substance from the mixture by filtration, thefiltrate was washed with ethyl acetate (50 ml.). Ethyl acetate (70 ml.)was added to the solution, and the mixture was adjusted to pH 1.5 with10% hydrochloric acid and then shaked sufficiently. After the ethylacetate layer was removed, the aqueous layer was extracted with ethylacetate (30 ml.) twice. The ethyl acetate layer and the extracts werecombined together, washed with a saturated aqueous solution of sodiumchloride, dried over magnesium sulfate and then concentrated underreduced pressure to give7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylicacid (syn isomer 0.48 g.), m.p. 165° to 174° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3250, 1780, 1730, 1690, 1660, 1550 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.57 (2H, broad s), 3.91 (3H, s), 5.30 (1H, d,J=5 Hz), 5.88 (1H, dd, J=5, 8 Hz), 7.44 (1H, s), 8.52 (1H, s), 9.78 (1H,d, J=8 Hz), 12.60 (1H, s).

(3)7-{2-(2-Formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 0.4 g.) was suspended in methanol (15 ml.). Afteradding conc-hydrochloric acid (0.16 g.) to the suspension, the mixturewas stirred at room temperature for 2.5 hours. Methanol was distilledoff from the resultant mixture under reduced pressure, and the residuewas dissolved in water (15 ml.). The solution was washed with ethylacetate (30 ml.) and dichloromethane (30 ml.) in turn. To the aqueouslayer was introduced nitrogen gas to remove the remaining organicsolvent completely, and the solution was lyophilized to give7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylicacid hydrochloride (syn isomer 0.35 g.), m.p. 170° to 180° C. (dec.)

I.R. ν_(max) ^(Nujol) : 3300, 1780, 1730, 1670, 1630, 1545 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.88 (2H, AB-q, J=17 Hz), 3.94 (3H, s), 5.26(1H, d, J=5 Hz), 5.80 (1H, dd, J=5, 8 Hz), 6.92 (1H, s), 9.88 (1H, d,J=8 Hz).

(4)7-[2-(2-Amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid hydrochloride (syn-isomer: 1.5 g.) and sodium bicarbonate (0.56 g.)were dissolved in water (50 ml.) at room temperature with stirring andlyophilized. A solution of iodomethyl hexanoate (0.93 g.) indimethylformamide (5 ml.) was added dropwise to a solution of theproduct obtained above in dimethylformamide (15 ml.) at -5° C. andstirred at the same temperature for 30 minutes. Ethyl acetate (50 ml.)and water (100 ml.) were added to the resultant solution and the ethylacetate layer was separated. The aqueous layer was extracted with ethylacetate (50 ml.) twice. The extracts were combined with the ethylacetate layer, washed with a saturated aqueous solution of sodiumbicarbonate three times and a saturated aqueous solution of sodiumchloride three times in turn, dried over magnesium sulfate, treated withactivated charcoal and then concentrated under reduced pressure. Afterwashing the concentrate with n-hexane (50 ml.), n-hexane (50 ml.) anddiethylether (25 ml.) were added to the residue and allowed to stand ina refrigerator overnight. The precipitating powder was collected byfiltration, washed with n-hexane and dried to give a mixture (1.0 g.) ofn-hexanoyloxymethyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn-isomer) and n-hexanoyloxymethyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-2-cephem-4-carboxylate(syn-isomer.)

(5) Thus obtained mixture (1.0 g.) was added to methylene chloride (10ml.) To the solution were added acetic acid (7 ml.), a solution ofsodium tungstate (Na₂ WO₄.2H₂ O) (20 mg.) in water (0.5 ml.), methylenechloride (5 ml.) and 35% hydrogen peroxide (180 mg.), and then stirredunder ice-cooling for 4 hours. Ice-water was added to the resultantsolution and extracted with methylene chloride. The extract was washedwith water, dried over magnesium sulfate and concentrated under reducedpressure. The residue was pulverized with diethyl ether (20 ml.), washedwith diethyl ether (10 ml.) twice and dried. The product was purifiedwith column chromatography on silica gel (eluent: ethyl acetate) to given-hexanoyloxymethyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate-1-oxide(syn-isomer: 600 mg.)

I.R. ν_(max) ^(Nujol) : 3300, 1790, 1760, 1680, 1630, 1540, 1380 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 0.67-2.5 (11H, m), 3.90 (3H, s), 4.20 (2H,broad s), 5.17 (1H, d), 5.83-6.17 (3H, m), 6.88 (1H, s), 9.17 (1H, d).

(6) Phosphorus trichloride (210 mg.) was added to a solution ofn-hexanoyloxymethyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate-1-oxide(syn-isomer: 570 mg.) in dry dimethylformamide (10 ml.) at -30° C., andstirred at -20° to -30° C. for 50 minutes. 10% Aqueous solution (50 ml.)of sodium chloride was added to the resultant solution, adjusted to pH8.0 with a saturated aqueous solution of sodium bicarbonate andextracted with ethyl acetate. The extract was washed with a saturatedaqueous solution of sodium bicarbonate and a saturated aqueous solutionof sodium chloride, dried over magnesium sulfate, and then concentratedunder reduced pressure. The residue (560 mg.) was purified with columnchromatography on silica gel (20 g.) (eluent: ethyl acetate), and theresultant residue (180 mg.) was pulverized with n-hexane (10 ml.) anddiethyl ether (5 ml.) to give n-hexanoyloxymethyl 7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn-isomer: 150 mg.).

I.R. ν_(max) ^(Nujol) : 3400 (broad), 1780 (broad), 1760 (shoulder),1670, 1620, 1530 cm⁻¹.

N.M.R. δ_(ppm) (CDCl₃): 0.67-2.5 (11H, m), 3.67 (2H, q), 4.00 (3H, s),5.17 (1H, d), 5.90 (1H, s), 6.00 (1H, m), 6.77 (1H, s), 7.83 (1H, d).

EXAMPLE 2

(1) 2-{2-(2,2,2-Trichloroacetamido)-4-thiazolyl}-2-methoxyiminoaceticacid (syn isomer, 0.65 g.) was added at 0° C. to Vilsmeier reagent whichhad been prepared from dimethylformamide and phosphorus oxychloride inethyl acetate (10 ml.), and the mixture was stirred at the sametemperature for 40 minutes to prepare the activated acid solution. Theactivated acid solution was added dropwise to a solution of7-amino-2,3-dimethyl-3-cephem-4-carboxylic acid (0.5 g.) andtrimethylsilylacetamide (1.73 g.) in ethyl acetate (30ml.) at -20° C.,and the mixture was stirred at the same temperature for 40 minutes. Tothe resultant mixture was added water (10 ml.), and the ethyl acetatelayer was separated from the mixture and washed with water. Water (30ml.) was added to the solution and the mixture was adjusted to pH 7.5with sodium bicarbonate under ice-cooling. After shaking the mixture,the aqueous layer was separated. Ethyl acetate (50 ml.) was added to theaqueous solution, and the mixture was adjusted to pH 2 with dilutehydrochloric acid with stirring, and the ethyl acetate layer wasseparated, washed with water and a saturated aqueous solution of sodiumchloride in turn, treated with activated charcoal, dried over magnesiumsulfate and then concentrated under reduced pressure. The residue waspulverized with diisopropyl ether to give7-[2-{2-(2,2,2-trifluoroacetamido)-4-thiazolyl}-2-methoxyiminoacetamido]-2,3-dimethyl-3-cephem-4-carboxylicacid (syn isomer, 0.9 g.).

I.R. ν_(max) ^(Nujol) : 3250, 1780, 1725, 1680, 1650 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 1.43 (3H, d, J=8 Hz), 1,92 (1H, s), 3.82 (3H,s), 3.98 (1H, q, J=8 Hz), 5.18 (1H, d, J=6 Hz), 5.73 (1H, AB-q, J=6 Hz),7.43 (1H, s), 9.63 (1H, d, J=8 Hz).

(2)7-[2-{2-(2,2,2-Trifluoroacetamido)-4-thiazolyl}-2-methoxyiminoacetamido]-2,3-dimethyl-3-cephem-4-carboxylicacid (syn isomer, 0.86 g.) was dissolved in an aqueous solution (9 ml.)containing sodium acetate trihydrate (2.3 g.), and the solution wasstirred at room temperature for 19 hours. After removing the insolublesubstance from the resultant mixture by filtration, the filtrate wasadjusted to around pH 2.5 with 10% hydrochloric acid under ice-cooling.The precipitates were collected by filtration, washed with water anddried to give7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-2,3-dimethyl-3-cephem-4-carboxylicacid (syn isomer, 0.16 g.).

I.R. ν_(max) ^(Nujol) : 3320, 3200, 1770 (shoulder), 1670, 1630 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 1.44 (3H, d, J=7 Hz), 1.98 (3H, s), 3.57 (1H,q, J=7 Hz), 3.82 (3H, s), 5.18 (1H, d, J=5 Hz), 5.73 (1H, dd, J=5, 8Hz), 6.76 (1H, s), 9.63 (1H, d, J=8 Hz).

EXAMPLE 3

(1) A mixture of2-{2-(2,2,2-trifluoroacetamido)-4-thiazolyl}-2-methoxyiminoacetic acid(syn isomer, 0.8 g.), dimethylformamide (0.20 g.), phosphorusoxychloride (0.41 g.) and ethyl acetate (10 ml.) was stirred for 30minutes under ice-cooling to prepare the activated acid solution in asimilar manner to that of Example 2-(1). On the other hand, a solutionof 7-amino-3-methoxy-3-cephem-4-carboxylic acid hydrochloride (0.6 g.)and trimethylsilylacetamide (3 g.) in ethyl acetate (15 ml.) was stirredat 40° C. for 3 hours. To the solution was added dropwise the activatedacid solution at -10° to -20° C. in 2 minutes, and the mixture wasstirred at the same temperature for 1.5 hours. After water (10 ml.) wasadded to the resultant mixture, the ethyl acetate layer was separatedand allowed to stand. The precipitates were collected by filtration togive 7-[2-{2-(2,2,2-trifluoroacetamido)-4-thiazolyl}-2-methoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer, 0.3 g.). The aqueous layer, which was separated fromthe ethyl acetate layer, was extracted with ethyl acetate, and theextract was combined with the mother liquor obtained above. The ethylacetate solution was washed with a saturated aqueous solution of sodiumchloride, dried over magnesium sulfate and then filtered. The filtratewas concentrated under reduced pressure, and the residue was washed withdiethyl ether to give the same object compound (0.25 g.), Total yield0.55 g.

I.R. ν_(max) ^(Nujol) : 3230, 1770, 1715, 1650, 1580 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.62 (2H, AB-q, J=16 Hz), 3.78 (3H, s), 3.93(3H, s), 5.17 (1H, d, J=4 Hz), 5.54 (1H, dd, J=8, 4 Hz), 7.60 (1H, s),9.69 (1H, d, J=8 Hz).

(2) A solution of7-[2-{2-(2,2,2-trifluoroacetamido)-4-thiazolyl}-2-methoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer, 0.55 g.) and sodium acetate trihydrate (1,76 g.) inethyl acetate (3 ml.), tetrahydrofuran (3 ml.) and water (5.5 ml.) wasstirred at room temperature overnight. The aqueous layer was separatedfrom the resultant mixture, washed with dichloromethane, and thenevaporated under reduced pressure to remove the organic solvent. Theaqueous solution was adjusted to pH 4.2 under ice-cooling, and subjectedto column chromatography on Diaion HP-20 resin (Trade mark: manufacturedby Mitsubishi Chemical Industries Ltd., 15 ml.). After washing thecolumn with water, the object compound was eluted with 20% aqueousisopropyl alcohol. The eluate was concentrated under reduced pressureand the residue was lyophilized to give7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-3-methoxy-3-cephem-4-carboxylicacid (syn isomer 0.4 g.), m.p. 185° to 190° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3300, 1770, 1660, 1630, 1540 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.60 (2H, broad s), 3.75 (3H, s), 3.83 (3H,s), 5.12 (1H, d, J=4 Hz), 5.55 (1H, dd, J=4, 8 Hz), 6.82 (1H, s), 9.52(1H, d, J=8 Hz).

EXAMPLE 4

(1) To a suspension of p-nitrobenzyl7-phenylacetamido-3-cephem-4-carboxylate (10.50 g.) in drydichloromethane (100 ml.) was added dry pyridine (2.14 g.). Phosphoruspentachloride (5.50 g.) was added to the solution at -10° C., and themixture was stirred at -5° C. for 45 minutes and further at 10° C. foran hour. After adding methanol (520 g.) to the resultant mixture, themixture was stirred at -20° C. for 1.5 hours. The precipitates werecollected by filtration, washed with dichloromethane (120 ml.) anddiethyl ether (130 ml.) in turn, and then dried to give p-nitrobenzyl7-amino-3-cephem-4-carboxylate (7.90 g.), m.p. 182° C. (dec.).

I.R. ν_(max) ^(Nujol) : 1790, 1730, 1638, 1600 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.78 (2H, d, J=4 Hz), 5.27 (2H, dd, J=5 Hz),5.44 (2H, s), 6.78 (1H, t, J=4 Hz), 7.72 (2H, d, J=9 Hz), 8.26 (2H, d,J=9 Hz).

(2) Vilsmeier reagent prepared from dimethylformamide (0.43 g.) andphosphorus oxychloride (0.92 g.) was suspended in dry ethyl acetate (10ml.). To the suspension was added2-(2-formamido-4-thiazolyl)-2-methoxyimino acetic acid (syn isomer, 1.15g.) under ice-cooling with stirring, and the mixture was stirred at thesame temperature for 30 minutes to prepare the activated acid solution.On the other hand, p-nitrobenzyl 7-amino-3-cephem-4-carboxylatehydrochloride (1.79 g.) and trimethylsilylacetamide (5.0 g.) weredissolved in ethyl acetate (40 ml.). To the solution was added theactivated acid solution at -20° C. all at once, and the mixture wasstirred at the same temperature for 2.5 hours. Water (60 ml.) and ethylacetate (200 ml.) were added to the resultant solution, and the ethylacetate layer was separated, washed with 10% hydrochloric acid (60 ml.),a saturated aqueous solution of sodium bicarbonate (60 ml.) and anaqueous solution of sodium chloride (50 ml.) in turn, dried overmagnesium sulfate, treated with activated charcoal, and then evaporatedunder reduced pressure. Diethyl ether was added to the residue, and theprecipitates were collected by filtration to give p-nitrobenzyl7-{2-(2-formamido-4-thiazolyl)-2-methoxyimino-acetamido}-3-cephem-4-carboxylate(syn isomer, 1.30 g.), m.p. 210° to 212° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3240, 1780, 1730, 1690, 1655 1605, 1550, 1520cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.65 (2H, broad s), 3.90 (3H, s), 5.20 (1H, d,J=5 Hz), 5.43 (2H, s), 5.95 (1H, q, J=5, 8 Hz), 6.68 (1H, t, J=4 Hz),7.42 (1H, s), 7.72 (2H, d, J=9 Hz), 8.28 (2H, d, J=9 Hz), 8.46 (1H, s),9.72 (1H, d, J=8 Hz).

(3) To a solution of p-nitrobenzyl7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylate(syn isomer, 1.25 g.) in methanol (40 ml.) and tetrahydrofuran (50 ml.)was added 10% palladium carbon (0.65 g.), and the mixture was subjectedto catalytic reduction at room temperature under atmospheric pressurefor 3.5 hours. After removing the catalyst from the reaction mixture,the filtrate was concentrated under reduced pressure. Water (80 ml.) wasadded to the residue, and the mixture was adjusted to pH 7.5 with anaqueous solution of sodium bicarbonate, and then the insoluble substancewas filtered off. The filtrate was washed with ethyl acetate (50 ml.),and then ethyl acetate (100 ml.) was added to the solution. Afteradjusting to pH 1.5 with 10% hydrochloric acid, the ethyl acetate layerwas separated. The remaining aqueous layer was extracted with ethylacetate (80 ml.) twice, and the extracts were combined with the ethylacetate layer obtained above, washed with an aqueous solution of sodiumchloride, dried over magnesium sulfate, and then concentrated underreduced pressure to give7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylicacid (syn isomer, 0.60 g.), m.p. 176° to 183° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3250, 1780, 1690, 1660, 1550 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.63 (2H, d, J=4 Hz), 3.93 (3H, s), 5.10 (1H,d, J=5 Hz), 5.90 (1H, q, J=5, 8 Hz), 6.53 (1H, t, J=4 Hz), 7.47 (1H, s),8.57 (1H, s), 9.70 (1H, d, J=8 Hz), 12.63 (1H, s).

(4)7-{2-(2-Formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylicacid (syn isomer, 95 mg.) was suspended in methanol (4 ml.).

To the suspension was added conc. hydrochloric acid (110 mg.) and thesolution was stirred at room temperature for 4 hours. After distillingmethanol under reduced pressure, the residue was dissolved in water (30ml.) and the aqueous solution was washed with ethyl acetate (10 ml.) anddichloromethane (15 ml.) in turn. Nitrogen gas was introduced into theaqueous solution to exclude the remaining organic solvent, and theaqueous solution was lyophilized to give7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylicacid hydrochloride (syn isomer, 83 mg.), m.p. 180° to 190° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3300, 1770, 1710, 1660, 1630 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.64 (2H, broad s), 3.95 (3H, s), 5.14 (1H, d,J=5 Hz), 5.82 (1H, t, J=4 Hz), 6.95 (1H, s), 9.80 (1H, d, J=8 Hz).

(5) The solution of7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylicacid (syn isomer 10.8 g.), conc. hydrochloric acid (11 g.) and methanol(350 ml.) was stirred at room temperature for 4 hours. Afterconcentrating the resultant solution under reduced pressure, ethylacetate was added to the residue. The solution was adjusted to pH 8.0with a saturated aqueous solution of sodium bicarbonate and the aqueouslayer was separated and washed with diethyl ether. After nitrogen gaswas bubbled in the aqueous solution, the aqueous solution was adjustedto pH 4.0 with 10% hydrochloric acid. The precipitates were collected byfiltration and washed with water to give7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylicacid (syn isomer, 8.2 g.), m.p.>290° C.

IR ν_(max) ^(Nujol) : 3470, 3280, 3200, 1780, 1695, 1655, 1622 cm⁻¹.

NMR δppm (DMSO-d₆): 3.60 (2H, broad s), 3.84 (3H, s), 5.12 (1H, dd, J=5Hz), 5.84 (1H, dd, J=5, 8 Hz), 6.52 (1H, broad t) 6.76 (1H, s), 7.26(2H, broad s), 9.65 (1H, d, J=8 Hz).

(6) Sodium bicarbonate (1.04 g.) was added to a solution of7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid hydrochloride (syn isomer, 2.6 g.) in water (100 ml.) underice-cooling and stirred at room temperature. The resultant solution waslyophilized to give sodium7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer).

I.R. ν_(max) ^(Nujol) : 3100, 1760, 1650, 1590, 1530 cm⁻¹.

N.M.R. δ (D₂ O, ppm) 3.60 (2H, broad q), 4.00 (3H, s), 5.22 (1H, d),5.88 (1H, d), 6.35 (1H, q), 7.03 (1H, s).

(7) The product obtained above was dissolved in dryN,N-dimethylformamide (20 ml.). To the solution was dropwise added asolution of iodomethyl n-hexanoate (1.33 g.) and dryN,N-dimethylformamide (5 ml.) at -40° C. over 5 minutes, and thenstirred at the same temperature for 40 minutes and then underice-cooling for 45 minutes. The resultant solution was added to a mixedsolution of ethyl acetate (60 ml.) and water (125 ml.). The ethylacetate layer was separated, washed with a saturated sodium bicarbonateaqueous solution and a staturated sodium chloride aqueous solution inturn dried over magnesium sulfate, and then treated with an activatedcharcoal. After removing ethyl acetate from the solution, the residuewas triturated with diethyl ether to give n-hexanoyloxymethyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 750 mg.)

I.R. ν_(max) ^(Nujol) : 3170, 1780, 1750 (shoulder), 1670, 1630, 1530cm⁻¹.

N.M.R. δ (CDCl₃, ppm) 0.68-1.84 (9H, m), 2.20-2.48 (2H, t), 3.20-3.80(2H, m), 4.02 (3H, s), 5.04 (1H, d), 5.60-6.20 (3H, m), 6.62 (1H, q),6.80 (1H, s), 7.72 (1H, d).

(8) p-Nitrobenzyl7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 1.1 g.) was suspended in a mixture of ethanol (10 ml.) andwater (15 ml.) 1N Aqueous solution of potassium hydroxide (6 ml.) wasadded dropwise to the suspension at 5° to 7° C. over 10 minutes andstirred for 10 minutes. The resultant solution was adjusted to pH 7.5with 10% hydrochloric acid, washed with ethyl acetate and adjusted to pH2.5 with 10% hydrochloric acid. The precipitating crystals werecollected by filtration to give the mixture of7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn-isomer: 0.32 g.) and 7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylic acid (syn-isomer: 0.035 g.)

(9)7-[2-(2-Amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn-isomer: 5 g.) was gradually added to an aqueous solution (30ml.) of sodium bicarbonate (1.04 g.) at 35° to 40° C., and stirred at50° to 53° C. for 30 minutes. After removing the insoluble substancefrom the resultant solution, the filtrate was treated with activatedcharcoal (0.3 g.), and filtered. The filtrate was lyophilized to givesodium7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 4.2 g.)

I.R. δ_(max) ^(Nujol) : 3300-3100, 1760, 1670, 1595, 1530 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.50 (2H, broad s), 3.83 (3H, s), 5.00 (1H, d,J=5 Hz), 5.68 (1H, dd, J=5 Hz, 8 Hz), 6.13 (1H, broad s), 6.73 (1H, s),7.3 (2H, broad s), 9.60 (1H, d, J=8 Hz).

(10)7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.15 g.) was added to an aqueous solution of calciumhydroxide (0.111 g.) in water (100 ml.), and the solution was stirred atroom temperature for 10 minutes. After the solution was filtered, thefiltrate was lyophilized to give calcium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 1.2 g.).

I.R. ν_(max) ^(Nujol) : 3350, 1760, 1670, 1590, 1535, 1465 cm⁻¹.

N.M.R. δ (D₂ O, ppm): 3.51 (1H, d, J=5 Hz), 3.59 (1H, d, J=3 Hz), 3.97(3H, s), 5.15 (1H, d, J=5 Hz), 5.82 (1H, d, J=5 Hz), 6.33 (1H, dd, J=5Hz, 3 Hz), 6.95 (1H, s).

(11)7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.15 g.) was added to a suspension of magnesiumhydroxide (0.088 g.) in water (100 ml.), and the mixture was stirred at70° C. for 30 minutes to give a solution. After the resultant solutionwas filtered, the filtrate was lyophilized to give magnesium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 1.1 g.).

I.R. ν_(max) ^(Nujol) : 3350, 1760, 1660, 1610, 1530, 1460 cm⁻¹.

N.M.R. δ (D₂ O, ppm): 3.53 (1H, d, J=5 Hz), 3.59 (1H, d, J=3 Hz), 3.96(3H, s), 5.16 (1H, d, J=5 Hz), 5.84 (1H, d, J=5 Hz), 6.32 (1H, dd, J=5Hz, 3 Hz), 7.98 (1H, s).

(12)7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.15 g.) was added to a solution of arginine (0.523g.) in water (50 ml.), and the solution was stirred at room temperaturefor 10 minutes.

After the resultant mixture was filtered, the filtrate was lyophilizedto give an arginine salt of7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.35 g.).

I.R. ν_(max) ^(Nujol) : 3350, 3150, 1770 1650 (broad), 1580, 1530, 1460cm⁻¹.

N.M.R. δ (D₂ O, ppm): 1.4-2.1 (4H, m), 3.22 (2H, t, J=6 Hz), 3.55 (1H,d, J=6 Hz), 3.65 (1H, d, J=3 Hz), 3.82 (1H, d, J=6 Hz), 3.97 (3H, s),5.18 (1H, d, J=5 Hz), 5.85 (1H, d, J=5 Hz), 6.33 (1H, dd, J=6 Hz; 3 Hz),7.00 (1H, s).

(13) Sodium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 1.21 g.) was added to a solution of lysine hydrochloride(0.55 g.) in water (12 ml.).

The solution was lyophilized to give a lysine salt of7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.6 g.).

I.R. ν_(max) ^(Nujol) : 3350, 3150, 1770, 1600 (broad), 1530, 1460 cm⁻¹.

N.M.R. δ (D₂ O, ppm): 1.3-2.1 (6H, m), 3.03 (2H, t, J=7 Hz), 3.54 (1H,d, J=5 Hz), 3.64 (1H, d, J=3 Hz), 3.80 (1H, d, J=6 Hz), 3.97 (3H, s),5.17 (1H, d, J=5 Hz), 5.84 (1H, d, J=5 Hz), 6.32 (1H, dd, J=5 Hz, 3Hz),6.99 (1H, s).

(14) 20% Aqueous solution of sodium hydroxide was added to a suspensionof7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 15 g.) in a mixture of ethanol (8 ml.) and water (8ml.) at room temperature to make a solution of pH 7.5. After filtrationand washing, the filtrate and washings were combined (which contained18.3 ml. of water) and added dropwise to ethanol (46 ml.) at 20° to 25°C. under stirring and stirred at the same temperature for 30 minutes.Ethanol (28 ml.) was added dropwise to the mixture over 30 minutes, andstirred at the same temperature for 2 hours. The precipitates werecollected by filtration, washed with ethanol (20 ml.) and dried in vacuoat room temperature to give plates of sodium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylatedihydrate (syn isomer, 13.5 g.), mp 260° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3430, 3250, 1760 (shoulder), 1745, 1650, 1630(shoulder), 1590, 1540 cm⁻¹.

(15) Sodium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 15 g.) was dissolved in water (13 ml.) at 35° to 45° C.under stirring. Warmed ethanol (52 ml., 30° C.) was added dropwise tothe stirred solution, and stirred at the same temperature for 5 minutesand then at room temperature for 2 hours. The precipitates werecollected by filtration, washed with ethanol and dried under reducedpressure to give plates of sodium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylatedihydrate (syn isomer, 13.45 g.).

(16) 4N Aqueous solution of sodium hydroxide was carefully addeddropwise to a stirred suspension of7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 52 g.) in water (100 ml.) below 5° C. to make asolution of pH 7.0 to 7.5. After filtration and washing, the combinedfiltrate and the washings (200 ml.) was added dropwise to ethanol (2 l.)under stirring over 30 minutes and stirred at room temperature for 15minutes and then at 5° to 10° C. for an hour. The precipitates werecollected by filtration, washed with ethanol (200 ml.) and dried invacuo at 30° C. to give amorphous sodium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 46.3 g.).

I.R. ν_(max) ^(Nujol) : 3400, 3300, 3170, 1750, 1650, 1580 cm⁻¹.

(17) A suspension of sodium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 10 g.) in methanol (250 ml.) was treated with a supersonicapparatus to make a clear solution. The solution was allowed to stand atroom temperature, and then stirred at the same temperature for 3 hours.The precipitates were collected by filtration and washed with methanolto give amorphous sodium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

(18) The crystals obtained in the above Example 4-(14) were dried overP₂ O₅ in vacuo for one day at room temperature to give another plates ofsodium7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer).

EXAMPLE 5

(1) Vilsmeier reagent prepared from dimethylformamide (0.22 g.) andphosphorus oxychloride (0.46 g.) was suspended in dry ethyl acetate (20ml.). 2-(2-Formamido-4-thiazolyl)-2-methoxyiminoacetic acid (antiisomer, 0.62 g.) was added to the suspension under ice-cooling withstirring, and the mixture was stirred at the same temperature for 30minutes to prepare the activated acid solution. The solution was addedall at once to a solution of p-nitrobenzyl7-amino-3-chloro-3-cephem-4-carboxylate (1 g.) andtrimethylsilylacetamide (2.58 g.) in ethyl acetate (20 ml.) at -20° C.with stirring, and the mixed solution was stirred at -10° to -20° C. for1.5 hours. To the resultant solution was added water (20 ml.), and thesolution was stirred at -20° C. After separating the ethyl acetatelayer, the aqueous layer was extracted with ethyl acetate (20 ml.). Theethyl acetate layer and the extract were combined together, washed with10% hydrochloric acid (20 ml.) twice, water (20 ml.) once, a 5% aqueoussolution of sodium bicarbonate (20 ml.) three times, and an aqueoussolution of sodium chloride (20 ml.) once in turn, dried, and thenconcentrated under reduced pressure. The residue was washed with diethylether (50 ml.) to give p-nitrobenzyl7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylate(anti isomer, 1.27 g.), m.p. 135° to 145° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3150 to 3300 (broad), 1780, 1730, 1670 to 1690(broad) cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.83 (2H, AB-q, J=17 Hz), 3.97 (3H, s), 5.23(1H, d, J=5 Hz), 5.41 (2H, s), 5.9 (1H, dd J=5,8 Hz), 7.62 (2H, d, J=8Hz), 8.0 (1H, s) 8.2 (2H, d, J=8 Hz), 8.42 (1H, s), 9.55 (1H, d, J=8Hz), 12.43 (1H, s).

(2) 10% Palladium carbon (0.6 g.) was added to a solution ofp-nitrobenzyl7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylate(anti isomer, 1.16 g.) in methanol (20 ml.) and tetrahydrofuran (40ml.), and the mixture was subjected to catalytic reduction at roomtemperature under atmospheric pressure for 5 hours. After removing thecatalyst from the reaction mixture, the filtrate was concentrated underreduced pressure. Water (30 ml.) and ethyl acetate (60 ml.) were addedto the residue, and the mixture was adjusted to pH 7.5 with an aqueoussolution of sodium bicarbonate and shaked sufficiently. The aqueouslayer was separated and ethyl acetate (90 ml.) was added to the aqueoussolution. The aqueous layer was adjusted to pH 2.5 with 10% hydrochloricacid with stirring under ice-cooling, and the ethyl acetate layer wasseparated. The remaining aqueous layer was extracted with ethyl acetate(30 ml.), and the extract and the ethyl acetate layer were combinedtogether, washed with an aqueous solution of sodium chloride, dried andthen concentrated under reduced pressure. The residue was washed withdiisopropyl ether to give7-{2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylicacid (anti isomer, 0.47 g.). The compound was colored at 210° C. anddecomposed at above than 250° C.

I.R. ν_(max) ^(Nujol) : 3250, 1780, 1720 (shoulder), 1670 to 1690 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.8 (2H, AB-q, J=17 Hz), 4.0 (3H, s), 5.21(1H, d, J=5 Hz), 5.83 (1H, dd, J=5,8 Hz), 8.05 (1H, s), 8.47 (1H, s),9.55 (1H, d, J=8 Hz), 12.55 (1H, broad s).

(3)7-{2-(2-Formamido-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylicacid (anti isomer, 0.4 g.) was suspended in methanol (15 ml). Conc.hydrochloric acid (0.16 g.) was added to the suspension and the mixturewas stirred at room temperature for 5 hours. The precipitates werecollected by filtration, washed with a mixed solvent of methanol anddiethyl ether (1:1), and dried to give7-{2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido}-3-chloro-3-cephem-4-carboxylicacid hydrochloride (anti isomer, 0.31 g.), m.p. above than 250° C.

I.R. ν_(max) ^(Nujol) : 3250, 3200, 1788, 1720, 1680, 1640 cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.81 (2H, AB-q, J=17 Hz), 4.08 (3H, s), 5.22(1H, d, J=5 Hz), 5.7 (1H, dd, J=5,8 Hz), 7.59 (1H, s), 9.5 (1H, d, J=8Hz).

EXAMPLE 6

(1) A solution of Vilsmeier reagent was prepared from drydimethylformamide (0.39 g.), dry ethyl acetate (1.2 ml.) and phosphorusoxychloride (0.84 g.) in a usual manner. To the solution was added asolution of 2-(1,2,3-thiadiazol-4-yl)-2-methoxyiminoacetic acid (synisomer, 0.93 g.) in ethyl acetate (10 ml.) at -15° C. to prepare theactivated acid solution. On the other hand, a p-nitrobenzyl7-amino-3-cephem-4-carboxylate (1.5 g.), trimethylsilylacetamide (4.6g.) and bis(trimethylsilyl)acetamide (1 ml.) in dry ethyl acetate (50ml.) was stirred at 45° C. for 5 hours to give a solution. To thesolution was added all at once the activated acid solution obtainedabove at -10° C. with stirring, and the mixed solution was stirred at-5° C. for 1.5 hours. Water was added to the reaction mixture, and theinsoluble product was separated by filtration, washed with ethyl acetateand water in turn, and then dried to give pale yellow powder ofp-nitrobenzyl7-{2-(1,2,3-thiadiazol-4-yl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylate(syn isomer, 1.9 g.), m.p. 243° to 245° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3250, 1782, 1725, 1655, 1630, 1600, 1520, 1345cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.69 (2H, AB-q, J=14 Hz), 4.00 (3H, s), 5.24(1H, d, J=5 Hz), 5.46 (2H, s), 6.00 (1H, dd, J=5,8 Hz), 6.68 (1H, t, J=4Hz), 7.7 to 8.4 (4H, m), 9.44 (1H, s), 9.88 (1H, d, J=8 Hz).

(2) 10% Palladium carbon (0.85 g.) was added to a solution ofp-nitrobenzyl7-{2-(1,2,3-thiadiazol-4-yl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylate(syn isomer, 1.65 g.) in methanol (70 ml.) and tetrahydrofuran (90 ml.),and the mixture was subjected to catalytic reduction at room temperatureunder atmospheric pressure for 3.5 hours. After removing the catalystfrom the reaction mixture by filtration, the filtrate was concentratedunder reduced pressure. Water was added to the residue, and the mixturewas adjusted to pH 7 to 8 with sodium bicarbonate, washed with ethylacetate, adjusted to pH 1.5 with 10% hydrochloric acid, and thenextracted with ethyl acetate. The extract was washed with a saturatedaqueous solution of sodium chloride, dried over magnesium sulfate, andthen filtered. The filtrate was concentrated under reduced pressure, andthe residue was pulverized with diethyl ether. The precipitates werecollected by filtration and the dried to give yellow powder of7-{2-(1,2,3-thiadiazol-4-yl)-2-methoxyiminoacetamido}-3-cephem-4-carboxylic acid (syn isomer, 0.3 g.), m.p. 200° to 210° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3250, 2550 to 2600, 1785, 1715, 1655, 1630, 1600cm⁻¹.

N.M.R. δ_(ppm) (DMSO-d₆): 3.58 (2H, AB-q, J=14 Hz), 4.00 (3H, s), 5.15(1H, d, J=5 Hz), 5.90 (1H, dd, J=5,8 Hz), 6.52 (1H, t, J=5 Hz), 9.38(1H, s), 9.84 (1H, d, J=8 Hz).

EXAMPLE 7

7-Amino-3-cephem-4-carboxylic acid (1.7 g.) and sodium bicarbonate (2.84g.) were dissolved in a mixture of water (35 ml.) and acetone (35 ml.).On the other hand, phosphorus oxychloride (1.95 ml.) was added dropwiseto a suspension of 2-(2-amino-4-thiazolyl)-2-methoxyiminoacetic acid(syn-isomer: 3.42 g.) in dry ethyl acetate (34 ml.) over 10 minutes at0° to 6° C., and the mixture was stirred at the same temperature for 30minutes. To the solution was added dropwise a solution oftrimethylsilylacetamide (2.39 g.) in ethyl acetate (5 ml.) at 0° to 6°C. over 20 minutes, and the mixture was stirred for 20 minutes. Afterphosphorus oxychloride (1.95 ml.) was added dropwise to the mixture atthe above temperature over 10 minutes, the mixture obtained thus wasstirred for 30 minutes. And further, dimethylformamide (1.29 ml.) wasadded dropwise to the mixture over 10 minutes at the same temperatureand stirred for one hour to give a clear solution. The solution wasadded dropwise to the solution of 7-amino-3-cephem-4-carboxylic acid at-5° to 5° C., over 30 minutes, at pH 6.5 to 7.5, and the reactionmixture was stirred for one hour at the same temperature. Ethyl acetate(200 ml.) was added to the resultant solution, and the aqueous layer wasseparated, washed with methylene chloride, bubbled with nitrogen gas andadjusted to pH 4 with acetic acid. The solution was subjected to columnchromatography on macroporous, non-ionic adsorption resin "Diaion HP-20"(Trade mark: manufactured by Mitsubishi Chemical Industries Ltd.) andeluted with 20% aqueous solution of isopropyl alcohol. The eluate wasconcentrated under reduced pressure and lyophilized to give7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer: 2.0 g.) The product was identified with the authenticsample by IR and NMR spectrum.

EXAMPLE 8

(1) Phosphorus oxychloride (1.2 g.) was added all at once to asuspension of 2-(2-amino-4-thiazolyl)-2-methoxyiminoacetic acid(syn-isomer: 1.23 g.) in ethyl acetate (12 ml.) at 5° C. and stirred at4° to 6° C. for 30 minutes. Trimethylsilylacetamide (1.0 g.) was addedto the solution and stirred at 4° to 6° C. for 30 minutes. Phosphorusoxychloride (1.2 g.) was added again to the solution and stirred for 15minutes. And further, dimethylformamide (0.5 g.) was added all at onceto the solution at 4° to 6° C. and stirred for 40 minutes to give aclear solution. On the other hand, p-nitrobenzyl7-amino-3-cephem-4-carboxylate hydrochloride (1.9 g.) was added to amixture of tetrahydrofuran (30 ml.) and acetone (10 ml.), and an aqueoussolution (20 ml.) of sodium bicarbonate (0.6 g.) was added to themixture. To the solution was added dropwise the solution obtained aboveat 0° to 5° C., pH 8.0. After stirring the mixture at -2° to 2° C., atpH 8.0 for 30 minutes, the insoluble substance was filtered out. Thefiltrate was extracted with ethyl acetate, and the extract was washedwith a saturated aqueous solution of sodium chloride, dried overmagnesium sulfate and then concentrated under reduced pressure. Theresidue was pulverized with diisopropyl ether to give p-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 1.6 g.)

I.R. δ_(max) ^(Nujol) : 3300, 1780, 1730, 1670, 1520 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.60 (2H, m), 3.81 (3H, s), 5.12 (1H, d, J=5 Hz),5.85 (1H, dd, J=5 Hz, 10 Hz), 6.64 (1H, m), 6.70 (1H, s), 7.20 (2H, s),7.65 (2H, d, J=10 Hz), 8.19 (2H, d, J=10 Hz), 9.60 (1H, d, J=10 Hz).

(2) p-Nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 7.8 g.) was suspended in a mixture of ethanol (60 ml.) andwater (60 ml.) 1N Aqueous solution of potassium hydroxide (45 ml.) wasadded dropwise to the stirred suspension under ice-cooling over 10minutes and stirred at 5° C. for 15 minutes. The resultant solution wasadjusted to pH 7.0 with conc. hydrochloric acid, washed with ethylacetate and the concentrated under reduced pressure to half of itsinitial volume. The concentrated solution was adjusted to pH 5.0 andsubjected to column chromatography on macroporous, non-ionic adsorptionresin "Diaion HP-20" (Trade mark; manufactured by Mitsubishi ChemicalIndustries Ltd.; 80 ml.), and eluted with 5% aqueous solution ofisopropyl alcohol. The fractions containing the object compound werecollected and adjusted to pH 3.2 with 10% hydrochloric acid. Theprecipitating crystals were collected by filtration, and dried to give7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn-isomer: 2.3 g.).

EXAMPLE 9

p-Nitrobenzyl 7-amino-3-cephem-4-carboxylate (3.4 g.) was suspended intetrahydrofuran (60 ml.) and an aqueous solution (20 ml.) of sodiumbicarbonate (1.2 g.) was added to the suspension. 1N Aqueous sodiumhydroxide (30 ml.) was added dropwise to the solution at 3° to 4° C. andstirred for 20 minutes. The resultant solution was adjusted to pH 7.0with 10% hydrochloric acid and concentrated under reduced pressure. Theinsoluble substance was filtered out and the filtrate was washed withethyl acetate. Acetone (30 ml.) was added to the filtrate and cooled to-5° C. A solution prepared from phosphorus oxychloride,dimethylformamide, trimethylsilylacetamide and2-(2-amino-4-thiazolyl)-2-methoxyiminoacetic acid (syn-isomer: 2.2 g.)in a similar manner to Example 7 was added to the solution obtainedabove at -5° to 0° C., at pH 7.5 to 8.5. The mixture was stirred at 3°to 7° C., at pH 7.5 to 8.5 for 2 hours, and the insoluble substance wasfiltered out. The aqueous layer was separated from the filtrate, washedwith ethyl acetate and adjusted to pH 3.0 to give7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn-isomer: 1.1 g.).

EXAMPLE 10

(1) Phosphoryl chloride (1.764 g.) was added to a suspension of2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetic acid (syn isomer, 1.0 g.) intetrahydrofuran (10 ml.) below 5° C. and stirred at the same temperaturefor 20 minutes. To the solution were added trimethylsilylacetamide (0.4g.) and N,N-dimethylformamide (0.4 g.), and the solution was stirredbelow 5° C. for 40 minutes [Solution A]. On the other hand,trimethylsilylacetamide (3.5 g.) was added to a suspension of4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (1.5 g.) in tetrahydrofuran(15 ml.), and stirred at room temperature for 1.5 hours. To the solutionwas added all at once the above Solution A at -20° C., and the solutionwas stirred at -5° to 0° C. for an hour. Water (20 ml.) was added to theresultant solution at -20° C., and the solution was adjusted to pH 7.5with an aqueous solution of sodium bicarbonate.

Tetrahydrofuran (70 ml.) and a saturated aqueous solution of sodiumchloride (50 ml.) were added to the solution, and the solution wasshaken sufficiently. The aqueous layer was separated and extracted withtetrahydrofuran. The tetrahydrofuran layer and extract were combined andwashed with a saturated aqueous solution of sodium chloride. Thesolution was dried over magnesium sulfate and concentrated under reducedpressure. The residue was triturated with diisopropyl ether to give4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 2.5 g.).

I. R. ν_(max) ^(Nujol) : 3330, 1780, 1730, 1680, 1640, 1610 cm⁻¹.

N.M.R. δppm(DMSO-d₆): 1.17 (3H, t, J=7 Hz), 3.50 (2H, m), 4.05 (2H, q,J=7 Hz), 5.10 (1H, d, J=5 Hz), 5.85 (1H, dd, J=5 Hz, 8 Hz), 6.67 (1H,s), 7.17 (2H, m), 7.63 (2H, d, J=8 Hz), 8.18 (2H, d, J=8 Hz), 10.13 (1H,d, J=8 Hz).

(2) Palladium on carbon (1.0 g.) moistened with water (3 ml.) was addedto a solution of 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 2.3 g.) in a mixture of tetrahydrofuran (30 ml.), methanol(15 ml.) and acetic acid (0.3 ml.), and the suspension was subjected tocatalytic reduction at room temperature under ordinary pressure for 2hours. After removing the catalyst from the resultant mixture byfiltration, the filtrate was concentrated under reduced pressure. Ethylacetate was added to the residue, and the solution was adjusted to pH7.5 with an aqueous solution of sodium bicarbonate. After removing theinsoluble substance by filtration, the aqueous solution was separated,washed with ethyl acetate, adjusted to pH 5.5 and then treated withactivated charcoal. The aqueous solution was adjusted to pH 3.2, and theprecipitates were collected by filtration and dried to give7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.6 g.).

I.R. ν_(max) ^(Nujol) : 3500, 3300, 3200, 1785, 1625, 1600 cm⁻¹.

N.M.R. δppm(DMSO-d₆): 1.20 (3H, t, J=7 Hz), 3.57 (2H, m), 4.08 (2H, q,J=7 Hz), 5.08 (1H, d, J=5 Hz), 5.83 (1H, dd, J=5 Hz, 8 Hz), 6.47 (1H,m), 6.73(1H, s), 7.20(2H, m), 9.58 (1H, d, J=8 Hz).

EXAMPLE 11

(1) Triethylamine (2.37 g.), dimethylaniline (7.12 g.) andtrimethylsilyl chloride (3.93 g.) were added to a stirred suspension of4-nitrobenzyl 7-(2-phenylacetamido)-3-hydroxy-3-cephem-4-carboxylate (10g.) in methylene chloride (200 ml.) in turn, and the solution wasstirred at room temperature for an hour. Phosphorus pentachloride (4.88g.) was added to the solution at -30° to -25° C. and stirred at -25° to-20° C. for 3 hours. Methanol (42 ml.) was added to the solution at -25°to -20° C., and stirred for an hour. To the solution was added water (35ml.) at -25° to -20° C., and the solution was stirred at roomtemperature. The precipitates were collected by filtration, washed withmethylene chloride and diethyl ether in turn, and dried to give4-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate (5.2 g.), mp 148°C. (dec.).

I.R. ν_(max) ^(Nujol) : 3440, 3300, 1760, 1740 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 2.8-3.7 (2H, m), 4.90 (1H, t, J=4 Hz), 5.29 (1H,d, J=4 Hz), 5.38 (2H, s), 7.71 (2H, d, J=8 Hz), 8.26 (2H, d, J=8 Hz).

(2) Phosphoryl chloride (2.87 g.) was dropwise added to a solution ofN,N-dimethylformamide (1.37 g.) in ethyl acetate (10 ml.) at 5° to 10°C. Ethyl acetate (40 ml.) was added to the solution, and stirred underice cooling for 40 minutes. To the solution was added2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetic acid (syn isomer, 3.58g.), and the solution was stirred at 0° to 5° C. for 40 minutes. Theresultant solution was added all at once to a mixture of 4-nitrobenzyl7-amino-3-hydroxy-3-cephem-4-carboxylate (5 g.), ethyl acetate (50 ml.),trimethylsilylacetamide (14.3 g.) and bis(trimethylsilyl)acetamide (5.8g.) at -15° C., and stirred at -20° to -15° C. for 1.2 hours. Water (50ml.) was added to the resultant solution at -25° to -20° C., and stirreduntil the temperature rise 5° C. The aqueous layer was separated andextracted with ethyl acetate. The ethyl acetate layer and extract werecombined, washed with a saturated aqueous solution of sodium chlorideand dried over magnesium sulfate. After the solution was concentrated toa volume of 50 ml. under reduced pressure, the precipitates werecollected by filtration and washed with ethyl acetate to give4-nitrobenzyl7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer, 3.5 g.), mp 163° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3210, 3160, 3050, 1780, 1665 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.0-4.2 (2H, m), 3.95 (3H, s), 5.28 (1H, d, J=4Hz), 5.41 (2H, s), 5.64 (1H, dd, J=4 Hz, 9 Hz), 7.49 (1H, s) 7.67 (2H,d, J=8 Hz), 8.21 (2H, d, J=8 Hz), 8.50 (1H, t, J=9 Hz).

(3) A solution of the compound obtained above (1 g.) in methanol (15ml.), tetrahydrofuran (5 ml.) and conc. hydrochloric acid (0.72 g.) wasstirred at room temperature for an hour. Diethyl ether (100 ml.) wasadded to the resultant solution and then triturated. The crystals werecollected by filtration to give 4-nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylatehydrochloride (syn isomer, 0.65 g.).

I.R. ν_(max) ^(Nujol) : 3180, 1780, 1680, 1670, 1640 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.2-4.0 (2H, m), 3.97 (3H, s), 5.27 (1H, d, J=4Hz), 5.41 (2H, s), 5.60 (1H, dd, J=4 Hz, 8 Hz), 7.10 (1H, s), 7.66 (2H,d, J=9 Hz), 8.25 (2H, d, J=9 Hz), 9.73 (1H, d, J=8 Hz).

EXAMPLE 12

(1) Phosphoryl chloride (1.76 g.) and trimethylsilylacetamide (0.4 g.)were added to a stirred suspension of2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetic acid (syn isomer, 1.0 g) intetrahydrofuran (10 ml.) below 5° C., and stirred at the sametemperature for 30 minutes. N,N-Dimethylformamide (0.4 g.) was added tothe solution and stirred below 5° C. for 20 minutes [Solution A].Trimethylsilylacetamide (4.8 g.) was added to a stirred suspension of4-nitrobenzyl 7-amino-3-chloro-3-cephem-4-carboxylate hydrochloride (1.9g.) in tetrahydrofuran (15 ml.), and the solution was stirred at roomtemperature for an hour. To the solution was added the above Solution Aall at once at -20° C., and the solution was stirred at 0° C. for anhour. Water (50 ml.) was added to the resultant solution at -20° C., andadjusted to pH 8.0 with an aqueous solution of sodium bicarbonate.Tetrahydrofuran (50 ml.) and a saturated aqueous solution of sodiumchloride (50 ml.) were added to the solution. The organic layer wasseparated, washed with a saturated aqueous solution of sodium chlorideand dried over magnesium sulfate. After concentrating the solution underreduced pressure, the residue was triturated with diisopropyl ether andthe precipitates were collected by filtration to give 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 2.0 g.).

I. R. ν_(max) ^(Nujol) : 3200, 1780, 1730, 1670 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 1.23 (3H, t, J=7 Hz), 3.96 (2H, s), 4.13 (2H, q,J=7 Hz), 5.31 (1H, d, J=5 Hz), 5.88 (1H, dd, J=5 Hz, 8 Hz), 6.77 (1H,s), 7.67 (2H, d, J=8 Hz), 8.25 (2H, d, J=8 Hz), 10.30 (1H, d, J=8 Hz).

(2) A suspension of palladium on carbon (0.8 g.) in water (5 ml.) wasadded to a mixture of 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 2.0 g.), acetic acid (0.6 ml.) and tetrahydrofuran (60ml.), and the suspension was subjected to catalytic reduction underordinary pressure at room temperature for 3 hours. After removing thecatalyst by filtration, the filtrate was concentrated under reducedpressure. After adding ethyl acetate (50 ml.) to the residue, thesolution was adjusted to pH 7.5 with an aqueous solution of sodiumbicarbonate, and the insoluble substance was filtered out. The aqueouslayer was separated and adjusted to pH 6.0 with 10% hydrochloric acid,and then the organic solvent was removed under reduced pressure. Theaqueous solution was subjected to column chromatography on macroporous,nonionic adsorption resin "Diaion HP-20" (Trademark, manufactured byMitsubishi Chemical Industries Ltd.) (30 ml.). The column was washedwith water and eluted with 5% aqueous isopropyl alcohol. The eluate waslyophilized to give sodium7-[2-(2-amino-4-thiazolyl)-2-ethoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 0.3 g.).

I.R. ν_(max) ^(Nujol) : 3350, 3200, 1770, 1675, 1620 cm⁻¹.

N.M.R. δppm (D₂ O): 1.33 (3H, t, J=7 Hz), 3.76 (2H, q, J=18 Hz, 30 Hz),4.30 (2H, q, J=7Hz), 5.33 (1H, d, J=5 Hz), 5.83 (1H, d, J=5 Hz), 7.06(1H, s).

EXAMPLE 13

(1) Phosphoryl chloride (4.6 g.), trimethylsilylacetamide (0.95 g.) andN,N-dimethylformamide (1.2 g.) were added to a stirred suspension of2-(2-amino-4-thiazolyl)-2-isopropoxyiminoacetic acid (syn isomer, 2.8g.) in tetrahydrofuran (25 ml.) below 5° C. for 30 minutes [Solution A].On the other hand, trimethylsilylacetamide (10.5 g.) was added to asuspension of 4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (3.9 g.) intetrahydrofuran (50 ml.), and stirred at room temperature for 1.5 hours.To the solution was added the above solution A at -20° C. all at once,and the solution was stirred at -5° to 0° C. for 40 minutes. Water (70ml.) and tetrahydrofuran (100 ml.) were added to the resultant solutionat -20° C. The solution was adjusted to pH 7.5 with an aqueous solutionof sodium bicarbonate and stirred for an hour. After a saturated aqueoussolution of sodium chloride (200 ml.) was added, the organic layer wasseparated. The remaining aqueous layer was extracted withtetrahydrofuran, and the extract and the above organic layer werecombined, washed with a saturated aqueous solution of sodium chloride,dried over magnesium sulfate and then concentrated under reducedpressure. The residue was triturated with diisopropyl ether and theprecipitates were collected by filtration to give 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-isopropoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 6.0 g.).

I.R. ν_(max) ^(Nujol) : 3320, 3270, 1775, 1730, 1670, 1630 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 1.17 (6H, d, J=6 Hz), 3.63 (2H, m), 4.33 (1H, q,J=6 Hz), 5.17 (1H, d, J=5 Hz), 5.42 (2H, s), 5.92 (1H, dd, J=5 Hz, 8Hz), 6.67 (1H, m), 6.70 (1H, s), 7.22 (2H, m), 7.70 (2H, d, J=8 Hz),8.25 (2H, d, J=8 Hz), 10.13 (1H, d, J=8 Hz).

(2) Acetic acid (1 ml.) and a suspension of 10% palladium on carbon (2.0g.) in water (8 ml) were added to a solution of 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-isopropoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 5.0 g.) in tetrahydrofuran (150 ml.), and the suspensionwas subjected to catalytic reduction at room temperature under ordinarypressure. After removing the catalyst by filtration, the filtrate wasconcentrated under reduced pressure. Ethylacetate (80 ml.) was added tothe residue, and adjusted to pH 7.5 with an aqueous solution of sodiumbicarbonate. The organic layer was separated and extracted with anaqueous solution of sodium bicarbonate. The extract and the aqueouslayer obtained above were combined, adjusted to pH 3.0 with conc.hydrochloric acid and extracted with tetrahydrofuran. The extract waswashed with a saturated aqueous solution of sodium chloride, dried overmagnesium sulfate and concentrated under reduced pressure.

The precipitating crystalls were collected by filtration and dried togive7-[2-(2-amino-4-thiazolyl)-2-isopropoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.8 g.).

I.R. ν_(max) ^(Nujol) : 3320, 1780, 1670, 1635 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 1.20 (6H, d, J=6 Hz), 3.55 (2H, m), 4.30 (1H, q,J=6 Hz), 5.08 (1H, d, J=5 Hz), 5.82 (1H, dd, J=5 Hz, 8 Hz), 6.45 (1H,m), 6.68 (1H, s), 7.10 (2H, m), 10.08 (1H, d, J=8 Hz).

EXAMPLE 14

(1) Phosphoryl chloride (4.6 g.), trimethylsilylacetamide (0.95 g.) andN,N-dimethylformamide (1.2 g.) were added to a stirred suspension of2-(2-amino-4-thiazolyl)-2-propoxyiminoacetic acid (syn isomer, 2.8 g.)in tetrahydrofuran (25 ml.) below 5° C., and stirred for 20 minutes. Thesolution was dropwise added to a suspension of 4-nitrobenzyl7-amino-3-cephem-4-carboxylate (3.9 g.) in a mixture of tetrahydrofuran(20 ml.), water (20 ml.) and acetone (20 ml.) at -5° to 5° C. whilekeeping the pH value at 6.9 to 7.1 with 20% aqueous solution of sodiumcarbonate. The solution was stirred at -5° to 5° C. for 30 minutes andfurther at 10° C. for an hour, and adjusted to pH 7.5. Aftertetrahydrofuran (100 ml.) and a saturated aqueous solution of sodiumchloride (200 ml.) were added to the resultant solution, the insolublesubstance was filtered out. The organic layer was separated from thefiltrate, washed with a saturated aqueous solution of sodium chloride,dried over magnesium sulfate and then concentrated under reducedpressure. The residue was triturated with diisopropyl ether and theprecipitates were collected by filtration to give 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-propoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 5.8 g.).

I.R. ν_(max) ^(Nujol) : 3300, 1780, 1730, 1670, 1640 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 0.93 (3H, t, J=6 Hz), 1.70 (2H, m), 3.70 (2H, m),4.08 (2H, t, J=6 Hz), 4.5 (2H, m), 5.23 (1H, d, J=5 Hz), 5.50 (2H, s),5.97 (1H, dd, J=5 Hz, 8 Hz), 6.73 (1H, m), 6.80 (1H, s), 7.75 (2H, d,J=9 Hz), 8.30 (2H, d, J=9 Hz), 9.65 (1H, d, J=8 Hz).

(2) 4-Nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-propoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 5.0 g.) was treated in a similar manner to that of Example13.(2) to give7-[2-(2-amino-4-thiazolyl)-2-propoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.9 g.)

I.R. ν_(max) ^(Nujol) : 3250, 1770, 1650, 1660, 1620 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 0.93 (3H, t, J=7 Hz), 1.67 (2H, sextet, J=7 Hz),3.60 (2H, m), 4.03 (2H, t, J=7 Hz), 5.13 (1H, d, J=5 Hz), 5.83 (1H, dd,J=5 Hz, 8 Hz), 6.48 (2H, t, J=4 Hz), 6.70 (1H, s), 7.18 (2H, m), 9.53(1H, d, J=8 Hz).

EXAMPLE 15

(1) Phosphoryl chloride (13.2 g.) was added dropwise to a stirredsolution of N,N-dimethylformamide (6.3 g.) and tetrahydrofuran (24.7ml.) at -5° C., and stirred at the same temperature for 30 minutes.Tetrahydrofuran (120 ml.) and2-(2-formamidothiazol-4-yl)-2-n-butoxyiminoacetic acid (syn isomer, 19.5g.) were added to the solution at -5° C., and stirred at the sametemperature for 30 minutes. The solution was added dropwise a stirredsuspension of 4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (24.7 g.),tetrahydrofuran (120 ml.), acetone (60 ml.) and water (60 ml.) at -5° to5° C. over 15 minutes while adjusting to pH 7 to 7.5 with 20% aqueoussolution of sodium carbonate, and then the solution was stirred for 30minutes. The insoluble substance was filtered off, and a saturatedaqueous solution of sodium chloride was added to the filtrate. Thesolution was extracted with tetrahydrofuran twice. The extract waswashed with a saturated aqueous solution of sodium chloride, dried overmagnesium sulfate and concentrated in vacuo. The residue was trituratedwith diisopropyl ether to give 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-n-butoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 34.6 g.).

I.R. ν_(max) ^(Nujol) : 3240, 3050, 1780, 1730, 1695, 1660 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.92 (3H, t, J=7 Hz), 0.8˜2.2 (4H, m), 3.67 (2H,d, J=4 Hz), 4.16 (2H, t, J=7 Hz), 5.23 (1H, d, J=5 Hz), 5.46 (2H, s),5.99 (1H, dd, J=5 Hz, 8 Hz), 6.71 (1H, t, J=5 Hz), 7.43 (1H, s), 7.76(2H, d, J=9 Hz), 8.30 (2H, d, J=9 Hz), 8.58 (1H, s), 9.72 (1H, d, J=8Hz), 12.66 (1H, s).

(2) A mixture of 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-n-butoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 34.5 g.), tetrahydrofuran (345 ml.), 10% palladium carbon(14 g.), methanol (140 ml.), acetic acid (2.5 ml.) and water (50 ml.)was subjected to catalytic reduction under ordinary pressure at roomtemperature for 3 hours. The resultant mixture was filtered, and washedwith tetrahydrofuran. The filtrate was concentrated in vacuo, and theresidue was dissolved in a mixture of ethyl acetate and an aqueoussolution of sodium bicarbonate. The insoluble substance was removed byfiltration. After the ethyl acetate layer was separated and extractedwith an aqueous solution of sodium bicarbonate, the aqueous layer andthe aqueous extract were combined. After the aqueous solution was washedwith ethyl acetate and diethyl ether in turn, the solution was adjustedto pH 2.0 with 10% hydrochloric acid and stirred for 30 minutes. Theprecipitates were collected by filtration, washed with water and driedover magnesium sulfate to give 7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 18.3 g.).

I.R. ν_(max) ^(Nujol) : 3330, 3040, 1780, 1725, 1695, 1655 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.90 (3H, t, J=7 Hz), 1.1˜1.9 (4H, m), 3.58 (2H,d, J=5 Hz), 4.12 (2H, t, J=7 Hz), 5.13 (1H, d, J=5 Hz), 5.86 (1H, dd,J=5 Hz, 8 Hz), 6.46 (1H, t, J=4 Hz), 7.40 (1H, s), 8.50 (1H, s), 9.63(1H, d, J=8 Hz), 12.57 (1H, broad s).

(3) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-n-butoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 12.7 g.), conc. hydrochloric acid (9.6 ml.), methanol(9.5 ml.) and tetrahydrofuran (9.5 ml.) was stirred at room temperaturefor 3 hours. The resultant solution was concentrated in vacuo, and theresidue was suspended in water. The suspension was adjusted to pH 3.5with sodium bicarbonate under ice cooling, and stirred at sametemperature for 30 minutes. The precipitates were collected byfiltration and dried over magnesium sulfate to give the powder (10 g.).The powder was suspended in water (300 ml.) and adjusted to pH 7.0 withsodium bicarbonate. The solution was adjusted to pH 6.0 with 10%hydrochloric acid and subjected to column chromatography on nonionicadsorption resin (Diaion HP-20: trademark, manufactured by MitsubishiChemical Industries Ltd.) (300 ml.) with 10% aqueous solution ofisopropyl alcohol. The eluate was adjusted to pH 3.5 with 10%hydrochloric acid under ice cooling, and the precipitates were collectedby filtration, washed with water and dried to give7-[2-(2-aminothiazol-4-yl)-2-n-butoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 7.2 g.).

I.R. ν_(max) ^(Nujol) : 3320, 1775, 1660 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.88 (3H, t, J=7 Hz), 1.1˜1.9 (4H, m), 3.58 (2H,broad s), 4.05 (2H, t, J=7 Hz), 5.08 (1H, d, J=5 Hz), 5.80 (1H, dd, J=5Hz, 8 Hz), 6.44 (1H, braod s), 7.18 (2H, s), 9.51 (1H, d, J=8 Hz).

EXAMPLE 16

(1) 2-(2-Formamidothiazol-4-yl)-2-iso-butoxyiminoacetic acid (synisomer, 6.48 g.), N,N-dimethylformamide (2.10 g.), phosphoryl chloride(4.40 g.), tetrahydrofuran (110 ml.), 4-nitrobenzyl7-amino-3-cephem-4-carboxylate (8.23 g.), acetone (16 ml.) and water (16ml.) were treated in a similar manner to that of Example 15-(1) to give4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-iso-butoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 12.8 g.).

I.R. ν_(max) ^(Nujol) : 3240, 3050, 1780, 1720, 1700, 1655 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.92 (6H, d, J=7 Hz), 1.7˜2.2 (1H, m), 3.67 (2H,broad s), 3.91 (2H, d, J=7 Hz), 5.21 (1H, d, J=5 Hz), 5.95 (1H, dd, J=5Hz, 9 Hz), 6.67 (1H, t, J=4 Hz), 7.37 (1H, s), 7.72 (2H, d, J=8 Hz),8.24 (2H, d, J=8 Hz), 8.52 (1H, s), 9.68 (1H, d, J=9 Hz), 12.58 (1H,broad s).

(2) 4-Nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-isobutoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 14.2 g.), 10% palladium carbon (5.7 g.), methanol (57 ml.),tetrahydrofuran (142 ml.), acetic acid (1 ml.) and water (10 ml.) weretreated in a similar manner to that of Example 15-(2) to give7-[2-(2-formamidothiazol-4-yl)-2-isobutoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 4.25 g.).

I.R. ν_(max) ^(Nujol) : 3260, 1790, 1725, 1670 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.92 (6H, d, J=6 Hz), 1.6˜2.3 (1H, m), 3.61 (2H,d, J=4 Hz), 3.91 (2H, d, J=6 Hz), 5.14 (1H, d, J=5 Hz), 5.88 (1H, dd,J=5 Hz, 8 Hz), 6.50 (1H, t, J=5 Hz), 7.40 (1H, s), 8.56 (1H, s), 9.64(1H, d, J=8 Hz).

(3)7-[2-(2-formamidothiazol-4-yl)-2-iso-butoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 4.1 g.), conc. hydrochloric acid (3.65 g.) andmethanol (61.5 ml.) were treated in a similar manner to that of Example15-(3) to give7-[2-(2-aminothiazol-4-yl)-2-iso-butoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 2.4 g.).

I.R. ν_(max) ^(Nujol) : 3330, 1780, 1665, 1630, 1545 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.89 (6H, d, J=7 Hz), 1.6˜2.2 (1H, m), 3.58 (2H,broad s), 3.84 (2H, d, J=7 Hz), 5.10 (1H, d, J=5 Hz), 5.82 (1H, dd, J=5Hz, 9 Hz), 6.46 (1H, braod s), 6.68 (1H, s), 7.20 (2H, s), 9.53 (1H, d,J=9 Hz).

EXAMPLE 17

(1) 2-(2-Formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetic acid (synisomer, 0.9 g.), N,N-dimethylformamide (266 mg.), phosphoryl chloride(557 mg.), tetrahydrofuran (20 ml.), 4-nitrobenzyl7-amino-3-cephem-4-carboxylate (1.05 g.), acetone (3 ml.) and water (3ml.) were treated in a similar manner to that of Example 15-(1) to give4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 1.69 g.).

I.R. ν_(max) ^(Nujol) : 3260, 3170, 3070, 1785, 1725, 1700, 1655 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.8˜2.2 (10H, m), 3.66 (2H, broad s), 4.10 (1H,m), 5.16 (1H, d, J=5 Hz), 5.42 (2H, s), 5.95 (1H, dd, J=5 Hz, 9 Hz),6.66 (1H, broad s), 7.37 (1H, s), 7.70 (2H, d, J=8 Hz), 8.22 (2H, d, J=8Hz), 8.50 (1H, s), 9.63 (1H, d, J=9 Hz), 12.60 (1H, broad s).

(2) 4-Nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 2.0 g.), 10% palladium carbon (0.8 g.), methanol (8 ml.),tetrahydrofuran (20 ml.), acetic acid (0.14 ml.) and water (1.4 ml.)were treated in a similar manner to that of Example 15-(2) to give7-[2-(2-formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.77 g.).

I.R. ν_(max) ^(Nujol) : 3275, 3070, 1780, 1675 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.8˜2.2 (10H, m), 3.62 (2H, broad s), 4.12 (1H,m), 5.13 (1H, d, J=5 Hz), 5.87 (1H, dd, J=5 Hz, 9 Hz), 6.47 (1H, broads), 7.37 (1H, s), 8.50 (1H, s), 9.58 (1H, d, J=9 Hz), 12.61 (1H, broads).

(3)7-[2-(2-Formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.72 g.), methanol (10.8 ml.) and conc. hydrochloricacid (0.61 g.) were treated in a similar manner to that of Example15-(3) to give7-[2-(2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.28 g.).

I.R. ν_(max) ^(Nujol) ; 3350, 1775, 1665, 1620, 1540 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.8˜2.2 (10H, m), 3.60 (2H, broad s), 4.04 (1H,m), 5.09 (1H, d, J=5 Hz), 5.83 (1H, dd, J=5 Hz, 9 Hz), 6.45 (1H, t, J=4Hz), 6.67 (1H, s), 7.19 (2H, s), 9.48 (1H, d, J=9 Hz).

EXAMPLE 18

Phosphoryl chloride (0.84 g.) was added dropwise to a stirred suspensionof 2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetic acid (syn isomer, 1.0g.), tetrahydrofuran (10 ml.) and water (0.05 ml.) at 5° C., and stirredat the same temperature for 20 minutes. Trimethylsilylacetamide (0.66g.), phosphoryl chloride (0.84 g.) and N,N-dimethylformamide (0.45 g.)were added to a solution, and stirred at 5° C. for an hour to preparethe activated acid solution. On the other hand, trimethylsilylacetamide(4.0 g.) was added to a suspension of 7-amino-3-cephem-4-carboxylic acid(0.88 g.) in tetrahydrofuran (10 ml.) at 40° C., and stirred for 30minutes. To the solution was added all at once the activated acidsolution obtained above at -20° C., and stirred at 0° C. for an hour.After water (20 ml.) was added to the resultant solution at -20° C., thesolution was adjusted to pH 7.5 with an aqueous solution of sodiumbicarbonate. Ethyl acetate was added to the solution, and the aqueouslayer was separated. The solution was washed with ethyl acetate anddiisopropyl ether in turn, adjusted to pH 5.0 and treated with activatedcharcoal. After the solution was adjusted to pH 3.0, the precipitateswere collected by filtration, washed with water and dried overphosphorus pentoxide to give7-[2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.8 g.).

I.R. ν_(max) ^(Nujol) : 3300, 1780, 1660, 1630 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.67 (2H, d, J=4 Hz), 4.67 (2H, m), 5.17 (1H, d,J=5 Hz), 5.25 (1H, m), 5.50 (1H, m), 5.90 (1H, dd, J=5 Hz, 8 Hz), 6.03(1H, m), 6.55 (1H, m), 6.80 (1H, s), 7.50 (2H, m), 9.68 (1H, d, J=8 Hz).

EXAMPLE 19

Phosphoryl chloride (1.4 g.) was added dropwise to a suspension of2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetic acid (syn isomer, 1.7g.) in tetrahydrofuran (15 ml.) below 7° C., and stirred at the sametemperature for 10 minutes. Phosphoryl chloride (1.4 g.),trimethylsilylacetamide (1.3 g.) and N,N-dimethylformamide (0.76 g.)were added to a solution, and stirred for 20 minutes to prepare theactivated acid solution. On the other hand, trimethylsilylacetamide (7.8g.) was added to a suspension of 7-amino-3-cephem-4-carboxylic acid (1.5g.) in tetrahydrofuran (20 ml.), and stirred at 40° C. for 30 minutes.To the solution was added all at once the activated acid solutionobtained above at -20° C., and stirred for 30 minutes at 0° C. Afteradding water (20 ml.) to the resultant solution at -20° C., the solutionwas adjusted to pH 7.5 with an aqueous solution of sodium bicarbonate.The aqueous layer was separated, washed with ethyl acetate anddiisopropyl ether in turn, and treated with activated charcoal at pH5.5. The solution was adjusted to pH 3.0, and the precipitates werecollected by filtration and dried over phosphorus pentoxide underreduced pressure to give7-[2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.47 g.).

I.R. ν_(max) ^(Nujol) : 3500, 3300, 1780, 1720, 1660, 1630 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.48 (1H, m), 3.67 (2H, m), 4.80 (2H, d, J=2Hz), 5.17 (1H, d, J=5 Hz), 5.88 (1H, dd, J=5 Hz, 8 Hz), 6.55 (1H, m),6.85 (1H, s), 7.33 (2H, m), 9.73 (1H, d, J=8 Hz).

EXAMPLE 20

(1) N,N-Dimethylformamide (3 drops) was added to a suspension of2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetic acid (syn isomer, 23g.) in thionyl chloride (230 ml.), and stirred at 60° C. for 5 minutes.After concentrating the solution in vacuo, benzene was added to theresidue. The presipitates were collected by filtration, washed withbenzene (30 ml.) three times and diethyl ether in turn to give2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetyl chloride (anti isomer,18 g.). On the other hand, trimethylsilylacetamide (46 g.) was added toa suspension of 4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (16.8 g.)in methylene chloride (168 ml.), and stirred at 40° C. for an hour. Tothe suspension was gradually added2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetyl chloride (anti isomer,13.6 g.) at -5° to 0° C., and stirred at the same temperature for anhour. Water (150 ml.) was added to the resultant solution and stirredfor 15 minutes. The recipitates were collected by filtration, washedwith water and dried over phosphorus pentoxide under reduced pressure togive 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(anti isomer, 25.7 g.).

I.R. ν_(max) ^(Nujol) : 3300 (broad), 1780, 1730, 1680, 1520 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.70 (2H, broad s), 4.07 (3H, s), 5.19 (1H, d,J=5 Hz), 9.57 (1H, d, J=8 Hz), 6.00 (1H, dd, J=5 Hz, 8 Hz), 5.30 (2H,s), 6.70 (1H, t, J=4 Hz), 7.71, 8.25 (4H, A₂ B₂, J=9 Hz), 8.07 (1H, s),8.50 (1H, s), 12.55 (1H, broad s).

(2) A suspension of 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(anti isomer, 4.2 g.), 10% palladium carbon (1.7 g.), acetic acid (0.63ml.), water (6.3 ml.), methanol (42 ml.), and tetrahydrofuran (84 ml.)was subjected to catalytic reduction in a hydrogen atmosphere at roomtemperature for 2 hours. After removing the catalyst by filtration, thefiltrate was concentrated to a volume of about 15 ml. under reducedpressure. Water (30 ml.) and ethyl acetate (50 ml.) were added to theconcentrated solution, and the solution was adjusted to pH 8.0 withsodium bicarbonate under stirring. The insoluble substance was removedby filtration, and the aqueous layer was separated and washed with ethylacetate (50 ml.). The solution was treated with activated charcoal, andadjusted to pH 2.2 with 10% hydrochloric acid under ice cooling. Theprecipitates were collected by filtration and washed with water to give7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (anti isomer, 2.52 g.).

I.R. ν_(max) ^(Nujol) : 3300 (broad), 1780, 1680, 1670, 1550 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.63 (2H, broad s), 4.08 (3H, s), 5.15 (1H, d,J=5 Hz), 5.87 (1H, dd, J=5 Hz, 8 Hz), 6.55 (1H, t, J=4 Hz), 8.09 (1H,s), 8.52 (1H, s), 9.46 (1H, d, J=8 Hz).

(3) A suspension of7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (anti isomer, 2.5 g.), conc. hydrochloric acid (2.5 ml.) andmethanol (38 ml.) was stirred at room temperature for two hours. Aftertreating the resultant solution with activated charcoal, the solutionwas concentrated in vacuo. The residue was crystallized out withdiisopropyl ether (100 ml.), and the precipitates were collected byfiltration, and washed with diisopropyl ether (30 ml.) to give7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid hydrochloride (anti isomer, 2.1 g.). The crystals were added towater (20 ml.) and adjusted to pH 6.0 with sodium bicarbonate. Thesolution was subjected to column chromatography on nonionic adsorptionresin "Diaion HP-20" (Trademark: manufactured by Mitsubishi ChemicalIndustries Ltd.] (75 ml.) with 10% diisopropyl ether. The eluate wasadjusted to pH 3.5 with 10% hydrochloric acid, and the precipitates werecollected by filtration and dried to give7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (anti isomer, 0.7 g.).

I.R. ν_(max) ^(Nujol) : 3400˜3200 (broad), 1770, 1680, 1640, 1520 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.60 (2H, d, J=5 Hz), 4.00 (3H, s), 5.10 (1H, d,J=5 Hz), 5.82 (1H, dd, J=5 Hz, 8 Hz), 6.48 (1H, t, J=4 Hz), 7.13 (2H,broad), 7.47 (1H, s), 9.42 (1H, d).

EXAMPLE 21

(1) The Vilsmeier reagent was prepared from N,N-dimethylformamide (0.4g.) and phosphoryl chloride (0.86 g.) in a usual manner. After thereagent was suspended in ethyl acetate (10 ml.),2-(2-formamidothiazol-4-yl)-2-n-propoxyiminoacetic acid (syn isomer, 1.3g.) was added to the stirred suspension under ice cooling and stirred atthe same temperature for 30 minute. The solution was added to a solutionof 4-nitrobenzyl 7-amino-3-chloro-3-cephem-4-carboxylate hydrochloride(2.0 g.), trimethylsilylacetamide (5.2 g.) and ethyl acetate (40 ml.) at-25° C., and stirred at -20° to -10° C. for 1.5 hours. After addingwater into the resultant solution, the solution was extracted with ethylacetate (60 ml.). The aqueous layer was extracted with ethyl acetate (50ml.). The extracts were combined together, washed with a saturatedaqueous solution of sodium bicarbonate and a saturated aqueous solutionof sodium chloride in turn, and dried over magnesium sulfate. Thesolution was concentrated in vacuo, and the residue was triturated withdiethyl ether. The precipitates were collected by filtratation, washedand dried to give 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-n-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 2.55 g.).

I.R. ν_(max) ^(Nujol) : 3250˜3150, 1780, 1730, 1690, 1660, 1610, 1550,1520 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.87 (3H, t, J=7 Hz), 1.63 (1H, m), 3.88 (2H, q,J=17 Hz), 3.97 (2H, q, J=7 Hz), 5.30 (1H, d, J=5 Hz), 5.40 (2H, s), 5.92(1H, dd, J=5 Hz, 8 Hz), 7.33 (1H, s), 7.65 (2H, d, J=9 Hz), 8.20 (2H, d,J=9 Hz), 8.47 (1H, s), 9.70 (1H, d, J=8 Hz), 12.40 (1H, s).

(2) A suspension of 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-n-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 2.4 g.), 10% palladium carbon (1.0 g.), methanol (24 ml.),water (3.6 ml.) and tetrahydrofuran (48 ml.) was subjected to catalyticreduction under ordinary pressure at room temperature. After removinginsoluble substance by filtration, the filtrate was concentrated invacuo. Water and ethyl acetate were added to the residue, and adjustedto pH 8 with a saturated aqueous solution of sodium bicarbonate. Theinsoluble substance was removed by filtration, and the aqueous layer wasseparated. Ethyl acetate was added to the solution, adjusted to pH 2.0with hydrochloric acid and extracted with ethyl acetate. The ethylacetate solution and extract were combined together, washed with asaturated aqueous solution of sodium chloride and dried over magnesiumsulfate. The solution was concentrated in vacuo, and the residue wastriturated with diethyl ether and collected by filtration to give7-[2-(2-formamidothiazol-4-yl)-2-n-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.6 g.).

I.R. ν_(max) ^(Nujol) : 3300˜3150, 1780, 1720, 1685, 1650, 1540 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.93 (3H, t, J=7 Hz), 1.72 (1H, m), 3.88 (2H, q,J=18 Hz), 4.08 (2H, q, J=7 Hz), 5.33 (1H, d, J=5 Hz), 5.92 (1H, dd, J=5Hz, 8Hz), 7.43 (1H, s), 8.57 (1H, s), 9.73 (1H, d, J=8 Hz).

(3) A suspension of7-[2-(2-formamidothiazol-4-yl)-2-n-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.5 g.), conc. hydrochloric acid (0.7 ml.) andmethanol (30 ml.) was stirred at room temperature for 1.5 hours. Afterremoving methanol from the resultant solution in vacuo, water (30 ml.)was added to the residue. After the solution was adjusted to pH 7.5 witha saturated aqueous solution of sodium bicarbonate, the insolublesubstance was removed by filtration. The filtrate was adjusted to pH 3with 10% hydrochloric acid. The precipitates were collected byfiltration and dried over phosphorus pentoxide to give7-[2-(2-aminothiazol-4-yl)-2-n-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 0.6 g.).

I.R. ν_(max) ^(Nujol) : 3300, 1780, 1670, 1630, 1530 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.92 (3H, t, J=7 Hz), 1.67 (1H, m), 3.70 (2H, q,J=18 Hz), 4.00 (2H, q, J=7 Hz), 5.25 (1H, d, J=5 Hz), 5.83 (1H, dd, J=5Hz, 8 Hz), 6.75 (1H, s), 9.63 (1H, d, J=8 Hz).

EXAMPLE 22

2-(2-Aminothiazol-4-yl)-2-n-hexyloxyiminoacetic acid (syn isomer, 3 g.),water (0.15 g.), phosphoryl chloride (3.8 g.), trimethylsilylacetamide(10.7 g.), N,N-dimethylformamide (1.0 g.), tetrahydrofuran (50 ml.) and7-amino-3-cephem-4-carboxylic acid (2.0 g.) were treated in a similarmanner to that of Example 18 to give7-[2-(2-aminothiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.1 g.).

I.R. ν_(max) ^(Nujol) : 3250, 1760, 1640, 1600 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.88 (3H, m), 1.1˜1.9 (8H, m), 3.60 (2H, m),4.06 (2H, t, J=6 Hz), 5.10 (1H, d, J=5 Hz), 5.82 (1H, dd, J=5 Hz, 8 Hz),6.46 (1H, m), 6.70 (1H, s), 7.26 (2H, m), 9.56 (1H, d, J=8 Hz).

EXAMPLE 23

2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetic acid (syn isomer,0.506 g.), N,N-dimethylformamide (0.151 g.), phosphoryl chloride (0.337g.) and ethyl acetate (7.9 ml.) were treated in a similar manner to thatof Example 15-(1) to give the activated acid solution.

On the other hand, trimethylsilylacetamide (1.85 g.) andbis(trimethylsilyl)acetamide (1.60 g.) were added to a suspension of4-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate (0.703 g.) inethyl acetate (10 ml.) and stirred at room temperature for an hour.

To the solution was added the activated acid solution obtained above at-10° C. all at once, and stirred at the same temperature for an hour.Water (20 ml.) and ethyl acetate (20 ml.) were added to the solution.The organic layer was separated, washed with a saturated aqueoussolution of sodium bicarbonate (8 ml.) and an aqueous solution of sodiumchloride (15 ml.), and dried over magnesium sulfate.

After the solution was concentrated in vacuo, the residue was pulverizedwith diethyl ether and the precipitates were collected by filtration togive 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer, 0.71 g.).

I.R. ν_(max) ^(Nujol) : 3270, 1770, 1740, 1670 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.39 (1H, m), 3.63 (2H, broad s), 4.90 (2H,broad s), 5.23-5.90 (4H, m), 7.57 (1H, s), 7.83 (2H, d, J=9 Hz), 8.40(2H, d, J=9 Hz), 8.67 (1H, s), 9.80 (1H, d, J=8 Hz), 12.83 (1H, broads).

EXAMPLE 24

2-(2-Formamidothiazol-4-yl)-2-propoxyiminoacetic acid (syn isomer, 0.515g.), N,N-dimethylformamide (0.161 g.), phosphoryl chloride (0.337 g.)and ethyl acetate (7.9 ml.) were treated in a similar manner to that ofExample 15-(1) to give the activated acid solution. On the other hand,trimethylsilylacetamide (1.85 g.) and bis(trimethylsilyl)acetamide (1.60g.) were added to a suspension of 4-nitrobenzyl7-amino-3-hydroxy-3-cephem-4-carboxylate (0.703 g.) in ethyl acetate (10ml.), and stirred at room temperature for an hour. To the solution wasadded the activated acid solution obtained above at -10° C. all at once,and stirred at the same temperature for an hour. To the resultantsolution were added water (20 ml.) and ethyl acetate (20 ml.). Theorganic layer was separated, washed with a saturated aqueous solution ofsodium bicarbonate (8 ml.) and water (14 ml.) and dried over magnesiumsulfate. After the solution was concentrated in vacuo, the residue waspulverized with diethyl ether, and the precipitates were collected byfiltration to give 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-propoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer, 0.75 g.).

I.R. ν_(max) ^(Nujol) : 3250, 1765, 1740, 1670 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.90 (3H, t, J=8 Hz), 1.67 (2H, m), 3.40 (2H,AB-q, J=20 Hz), 4.08 (2H, q, J=8 Hz), 5.03-5.83 (4H, m), 7.40 (1H, s),7.70 (2H, d, J=9 Hz), 8.27 (2H, d, J=9 Hz), 8.53 (1H, s), 9.50 (1H, d,J=8 Hz), 12.60 (1H, broad s).

EXAMPLE 25

2-(2-Formamidothiazol-4-yl)-2-isobutoxyiminoacetic acid (syn isomer,0.54 g.), N,N-dimethylformamide (0.16 g.), phosphoryl chloride (0.34 g.)and ethyl acetate (10 ml.) were treated in a similar manner to that ofExample 15-(1) to give the activated acid solution. On the other hand,trimethylsilylacetamide (1.85 g.) and bis(trimethylsilyl)acetamide (1.62g.) were added to a suspension of 4-nitrobenzyl7-amino-3-hydroxy-3-cephem-4-carboxylate (0.7 g.) in ethyl acetate (10ml.) and stirred at 40° C. for 30 minutes. The activated acid solutionobtained above was added to the solution at -20° C. all at once, andstirred at the same temperature for an hour. Water (10 ml.) was added tothe resultant solution, and the organic layer was separated, washed withwater, an aqueous solution of sodium bicarbonate and a saturated aqueoussolution of sodium chloride, and then dried. After the solution wasconcentrated in vacuo, the residue was pulverized with diisopropylether. The precipitates were collected by filtration, washed withdiisopropyl ether and dried to give 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-isobutoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer, 1.09 g.).

I.R. ν_(max) ^(Nujol) : 3250, 3050, 1750, 1650, 1610 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.97 (6H, d, J=6 Hz), 2.0 (1H, m), 3.60 (2H,AB-q, J=18 Hz), 3.95 (2H, d, J=6 Hz), 5.1-5.8 (4H, m), 7.53 (1H, s),7.63 (2H, d, J=8 Hz), 8.18 (2H, d, J=8 Hz), 8.58 (1H, s), 9.47 (1H, d,J=9 Hz), 12.77 (1H, broad s).

EXAMPLE 26

N,N-Dimethylformamide (0.114 g.), phosphoryl chloride (0.240 g.) andethyl acetate (0.5 ml.) were reacted in a conventional manner to give aVilsmeier reagent. Ethyl acetate (5 ml.) and2-(2-formamidothiazol-4-yl)-2-ethoxyiminoacetic acid (syn isomer, 0.35g.) were added at the same temperature for 30 minutes to give theactivated acid solution. On the other hand, trimethylsilylacetamide(1.31 g.) and bis(trimethylsilyl)acetamide (1.14 g.) were added to asuspension of 4-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate(0.50 g.) in ethyl acetate (7.5 ml.) and stirred at room temperature foran hour. To the solution was added the activated acid solution obtainedabove all at once at -10° C., and stirred at the same temperature for 30minutes. Water (15 ml.) and ethyl acetate (15 ml.) were added to theresultant solution. The organic layer was separated, washed with asaturated aqueous solution of sodium bicarbonate (15 ml.) and water (10ml.), dried over magnesium sulfate and concentrated in vacuo. Theresidue was pulverized with diethyl ether and the precipitates werecollected by filtration to give 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-ethoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer, 0.634 g.).

I.R. ν_(max) ^(Nujol) : 3220, 1760, 1740, 1670 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.24 (3H, t, J=8 Hz), 3.36 (2H, AB-q, J=20 Hz),4.15 (2H, q, J=8 Hz), 5.10-5.60 (4H, m), 7.37 (1H, s), 7.65 (2H, d, J=9Hz), 8.22 (2H, d, J=9 Hz), 8.48 (1H, s), 9.52 (1H, d, J=8 Hz), 12.58(1H, broad s).

EXAMPLE 27

(1) 2-(2-Formamidothiazol-4-yl)-2-pentyloxyiminoacetic acid (syn isomer,4.14 g.), 4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (4.5 g.),N,N-dimethylformamide (1.41 g.), phosphoryl chloride (2.96 g.),tetrahydrofuran (72 ml.), acetone (15 ml.) and water (15 ml.) weretreated in a similar manner to that of Example 15-(1) to give b4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-pentyloxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 8.1 g.).

I.R. ν_(max) ^(Nujol) : 3240, 3050, 1780, 1730, 1655 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6-2.0 (9H, m), 3.66 (2H, s), 4.10 (2H, t, J=6Hz), 5.19 (1H, d, J=5 Hz), 5.42 (2H, s), 5.95 (1H, dd, J=5 Hz, 8 Hz),6.16 (1H, broad s), 7.38 (1H, s), 7.72 (2H, d, J=9 Hz), 8.26 (2H, d, J=9Hz), 8.54 (1H, s), 9.69 (1H, d, J=8 Hz), 12.69 (1H, broad s).

(2) 4-Nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-pentyloxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 8 g.), 10% palladium carbon (3.6 g.), methanol (36 ml.)tetrahydrofuran (90 ml.), acetic acid (0.63 g.) and water (6.3 ml.) weretreated in a similar manner to that of Example 15-(2) to give7-[2-(2-formamidothiazol-4-yl)-2-pentyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 3.4 g.).

I.R. ν_(max) ^(Nujol) : 3275, 3075, 1795, 1700, 1660, 1630 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6-2.0 (9H, m), 3.60 (2H, d, J=4 Hz), 4.12 (2H,t, J=6 Hz), 5.14 (1H, d, J=5 Hz), 5.87 (1H, dd, J=5 Hz, 9 Hz), 6.49 (1H,t, J=3 Hz), 7.40 (1H, s), 8.53 (1H, s), 9.64 (1H, d, J=9 Hz), 12.68 (1H,s).

(3)7-[2-(2-Formamidothiazol-4-yl)-2-pentyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 3.3 g.), conc.hydrochloric acid (2.80 g.),tetrahydrofuran (20 ml.) and methanol (50 ml.) were treated in a similarmanner to that of Example 15-(3) to give7-[2-(2-aminothiazol-4-yl)-2-pentyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 2.3 g.)

I.R. ν_(max) ^(Nujol) : 3300, 1775, 1650, 1540 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6-2.0 (9H, m), 3.56 (2H, d, J=2 Hz), 4.03 (2H,t, J=6 Hz), 5.08 (1H, d, J=5 Hz), 5.81 (1H, dd, J=5 Hz, 8 Hz), 6.46 (1H,t, J=4 Hz), 6.69 (1H, s), 7.20 (2H, s), 9.15 (1H, d, J=8 Hz).

EXAMPLE 28

(1) Triethylamine (3.41 g.), N,N-dimethylphenylamine (10.3 g.) andtrimethylchlorosilan (5.64 g.) were added to a stirred solution of7-phenylacetamido-3-tosyloxy-3-cephem-4-carboxylic acid (15 g.) inmethylene chloride (150 ml.) at room temperature in turn, and thesolution was stirred at room temperature for an hour. Phosphoruspentachloride (7.03 g.) was added to the solution at -35° C., andstirred at -25° to -20° C. for 1.5 hours. Methanol (61 ml.) was added tothe solution and stirred at the same temperature for 40 minutes. To theresultant solution was added water (50 ml.) at -20° to -10° C. Theorganic layer was separated and washed with water twice. The aqueouslayer and the washings were combined, and washed with methylene chloridetwice and diethyl ether in turn. After the solution was adjusted to pH4.7 with 10% aqueous sodium hydroxide under cooling, the precipitateswere collected by filtration, washed with water, acetone and diethylether in turn, and then dried over phorphorus pentoxide to give7-amino-3-tosyloxy-3-cephem-4-carboxylic acid (5.01 g.), mp 172° C.(dec.).

I.R. ν_(max) ^(Nujol) : 3210, 1800, 1635, 1620 cm⁻¹.

N.M.R. δ(D₂ O+NaHCO₃, ppm): 2.45 (3H, s), 3.51 (2H, q, J=18 Hz), 5.08(1H, d, J=5 Hz), 5.51 (1H, d, J=5 Hz), 7.48 (2H, d, J=9 Hz), 7.84 (2H,d, J=9 Hz).

(2) 2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetic acid (syn isomer, 0.76g.), N,N-dimethylformamide (0.33 g.), phosphoryl chloride (1.46 g.),trimethylsilylacetamide (0.5 g.) and ethyl acetate (10 ml.) were treatedto give the activated acid solution in a conventional manner.

On the other hand, trimethylsilylacetamide (2.7 g.) was added to asuspension of 7-amino-3-tosyloxy-3-cephem-4-carboxylic acid (1.0 g.) inethyl acetate (15 ml.) and stirred at room temperature. To the solutionwas added the activated acid solution obtained above at -15° C. all atonce, and stirred at -5°--5° C. for an hour. After the resultantsolution was chilled to -20° C., water (30 ml.) was added to the chilledsolution and adjusted to pH 6.5 with an aqueous solution of sodiumbicarbonate. The insoluble substance was removed by filtration. Theaqueous layer was separated and adjusted to pH 3.0 with hydrochloricacid. The precipitate were collected by filtration and dried to give7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-tosyloxy-3-cephem-4-carboxylicacid (syn isomer, 1.0 g.).

I.R. ν_(max) ^(Nujol) : 3350, 1780, 1670, 1630 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 2.17 (3H, s), 3.71 (2H, m), 3.92 (3H, s), 5.32(1H, d, J=5 Hz), 5.87 (1H, dd, J=5 Hz, 8 Hz), 6.82 (1H, s), 7.50 (2H, d,J=8 Hz), 7.92 (2H, d, J=8 Hz), 9.73 (1H, d, J=8 Hz).

EXAMPLE 29

(1) 2-(2-Formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetic acid(syn isomer, 1.35 g.), 4-nitrobenzyl 7-amino-3-cephem-4-carboxylate(1.54 g.), N,N-dimethylformamide (393 mg.), phosphoryl chloride (825mg.), tetrahydrofuran (21.2 ml.), acetone (3.9 ml.) and water (3.9 ml.)were treated in a similar manner to that of Example 15-(1) to give4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer 2.52 g.).

I.R. ν_(max) ^(Nujol) : 3250, 1790, 1730, 1690, 1640 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.23 (3H, t, J=7 Hz), 3.66 (2H, s), 4.13 (2H, q,J=7 Hz), 4.74 (2H, s), 5.22 (1H, d, J=5 Hz), 5.42 (2H, s), 5.98 (1H, dd,J=5 Hz, 9 Hz), 6.49 (1H, broad s), 7.43 (1H, s), 7.71 (2H, d, J=9 Hz),8.23 (2H, d, J=9 Hz), 8.52 (1H, s), 9.68 (1H, d, J=9 Hz), 12.66 (1H, s).

(2) 4-Nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 2.52 g.), 10% palladium carbon (1.3 g.), ethanol (13 ml.),tetrahydrofuran (25 ml.), acetic acid (0.22 ml.) and water (2.2 ml.)were treated in a similar manner to that of Example 15-(2) to give7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.4 g.).

I.R. ν_(max) ^(Nujol) : 3250, 3060, 1780, 1750, 1690, 1660 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.23 (3H, t, J=7 Hz), 3.61 (2H, broad s), 4.15(2H, q, J=7 Hz), 4.73 (2H, s), 5.13 (1H, d, J=5 Hz), 5.87 (1H, dd, J=5Hz, 9 Hz), 6.48 (1H, broad s), 7.43 (1H, s), 8.50 (1H, s), 9.62 (1H, d,J=9 Hz), 12.58 (1H, s).

(3) A solution of7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.35 g.), conc.hydrochloric acid (0.39 g.), ethanol(5.3 ml.) and tetrahydrofuran (8 ml.) was stirred at room temperaturefor 4.5 hours. After the resultant solution was concentrated in vacuo,the residue was dissolved in an aqueous solution of sodium bicarbonate,treated with activated charcoal and filtered. The filtrate was adjustedto pH 3.5 with 10% hydrochloric acid under ice cooling.

The precipitates were collected by filtration, washed with water anddried to give7-[2-(2-aminothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.1 g.).

I.R. ν_(max) ^(Nujol) : 3250, 3050, 1775, 1720, 1660, 1630, 1550 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.21 (3H, t, J=7 Hz), 3.59 (2H, s), 4.14 (2H, q,J=7 Hz), 4.66 (2H, s), 5.10 (1H, d, J=5 Hz), 5.83 (1H, dd, J=5 Hz, 8Hz), 6.47 (1H, broad s), 6.78 (1H, s), 7.23 (2H, s), 9.52 (1H, d, J=8Hz).

EXAMPLE 30

(1) The Vilsmeier reagent was prepared from N,N-dimethylformamide (0.32g.) and phosphoryl chloride (0.67 g.) in a conventional manner. Afterthe reagent was suspended in ethyl acetate (10 ml.),2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxy)iminoacetic acid(syn isomer, 1.2 g.) was added to the stirred suspension under icecooling and stirred at the same temperature for 30 minutes. The solutionwas added to a solution of 7-amino-3-cephem-4-carboxylic acid (0.8 g.),and trimethylsilylacetamide (4.2 g.) in ethyl acetate (20 ml.) at -25°C., and stirred at -20° to -10° C. for an hour. Water and ethyl acetatewere added to the resultant solution, and ethyl acetate layer wasseparated. The aqueous layer was extracted again with ethyl acetate.Water was added to the combined extract and adjusted to pH 7.5 with asaturated aqueous solution of sodium bicarbonate, and then the aqueouslayer was separated. Ethyl acetate was added to the aqueous layer,adjusted to pH 1.5 with hydrochloric acid and the ethyl acetate layerwas separated. The aqueous layer was extracted again with ethyl acetate.The extracts were combined, washed with a saturated aqueous solution ofsodium chloride and dried over magnesium sulfate. After concentratingthe solution in vacuo, the residue was triturated with diethyl ether,and the precipitates were collected by filtration and dried to give7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.55 g.).

I.R. ν_(max) ^(Nujol) : 3250, 1790, 1690, 1660, 1630, 1605, 1580, 1550cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.67 (2H, broad s), 4.78 (2H, q, J=8.5 Hz), 5.17(1H, d, J=5 Hz), 5.92 (1H, dd, J=5 Hz, 8 Hz), 6.53 (1H, t, J=4 Hz), 7.52(1H, s), 8.57 (1H, s), 9.83 (1H, d, J=8 Hz), 12.67 (1H, broad s).

(2) A suspension of7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.5 g.), conc. hydrochloric acid (1.3 ml.),tetrahydrofuran (10 ml.) and methanol (30 ml.) was treated in a similarmanner to that of Example 21-(3) to give7-[2-(2-aminothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.1 g.).

I.R. ν_(mad) ^(Nujol) : 3450, 3300, 1780, 1660, 1625, 1590, 1550 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.60 (2H, broad s), 4.70 (2H, q, J=8.5 Hz), 5.13(1H, d, J=5 Hz), 5.87 (1H, dd, J=5 Hz, 8 Hz), 6.52 (1H, t, J=4 Hz), 6.87(1H, s), 9.80 (1H, d, J=8 Hz).

EXAMPLE 31

(1) A solution of2-(2-formamidothiazol-4-yl)-2-(2-chloroethoxyimino)acetic acid (synisomer, 3.47 g.), N,N-dimethylformamide (1.1 g.) and phosphoryl chloride(2.3 g.) in ethyl acetate (35 ml.), and a solution of7-amino-3-cephem-4-carboxylic acid (2.5 g.) andbis(trimethylsilyl)acetamide (12.7 g.) in ethyl acetate (25 ml.) weretreated in a similar manner to that of Example 15-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-(2-chloroethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.85 g.).

I.R. ν_(max) ^(Nujol) : 3250, 3050, 1780, 1695, 1685, 1655, 1625 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.62 (2H, d, J=4 Hz), 3.86 (2H, t, J=6 Hz), 4.37(2H, t, J=6 Hz), 5.16 (1H, d, J=5 Hz), 5.90 (1H, dd, J=5 Hz, 9 Hz), 6.52(1H, t, J=4 Hz), 7.50 (1H, s), 8.53 (1H, s), 9.68 (1H, d, J=9 Hz), 12.72(1H, broad s).

(2)7-[2-(2-Formamidothiazol-4-yl)-2-(2-chloroethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.8 g.), conc. hydrochloric acid (1.6 g.), methanol(27 ml.) and tetrahydrofuran (40 ml.) were treated in a similar mannerto that of Example 15-(3) to give7-[2-(2-aminothiazol-4-yl)-2-(2-chloroethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.4 g.).

I.R. ν_(max) ^(Nujol) : 3440, 3300, 3070, 1780, 1660, 1625, 1555 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.60 (2H, s), 3.80 (2H, t, J=6 Hz), 4.30 (2H, t,J=6 Hz), 5.10 (1H, d, J=5 Hz), 5.83 (1H, dd, J=5 Hz, 9 Hz), 6.47 (1H,s), 6.78 (1H, s), 7.24 (2H, s), 9.58 (1H, d, J=9 Hz).

EXAMPLE 32

(1) A solution of2-(2-formamidothiazol-4-yl)-2-(tert-butoxycarbonylmethoxyimino)aceticacid (syn isomer, 3.2 g.), N,N-dimethylformamide (0.852 g.) andphosphoryl chloride (1.79 g.) in ethyl acetate (34 ml.) and a solutionof 7-amino-3-cephem-4-carboxylic acid (1.95 g.) andbis(trimethylsilyl)acetamide (9.9 g.) in ethyl acetate (19.5 ml.) weretreated in a similar manner to that of Example 15-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-(tert-butoxycarbonylmethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer, 2.9 g.).

I.R. ν_(max) ^(Nujol) : 3260, 3180, 3060, 1785, 1730, 1690, 1640 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.44 (9H, s), 3.63 (2H, s), 4.62 (2H, s), 5.12(1H, d, J=5 Hz), 5.87 (1H, dd, J=5 Hz, 9 Hz), 6.48 (1H, broad s), 7.42(1H, s), 8.50 (1H, s), 9.57 (1H, d, J=9 Hz), 12.62 (1H, broad s).

(2) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-(tert-butoxycarbonylmethoxyimino)acetamido]-3-cephem-4-carboxylicacid (syn isomer, 2.8 g.), anisole (2.8 ml.) and trifluoroacetic acid(11.2 ml.) was stirred at room temperature for an hour. Ethyl acetateand water were added to the resultant solution and adjusted to pH 7.0with sodium bicarbonate. The aqueous layer was separated, and the ethylacetate layer was extracted with water. The aqueous extracts werecombined, washed with ethyl acetate and diethyl ether in turn, and thenadjusted to pH 2.0 with 10% hydrochloric acid under ice cooling. Theprecipitates were collected by filtration, washed with water and driedto give7-[2-(2-formamidothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.43 g.).

I.R. ν_(max) ^(Nujol) : 3270, 3120, 3070, 1760, 1720, 1690, 1660, 1620cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.60 (2H, s), 4.63 (2H, s), 5.11 (1H, d, J=5Hz), 5.88 (1H, dd, J=5 Hz, 9 Hz), 6.48 (1H, t, J=4 Hz), 7.44 (1H, s),8.52 (1H, s), 9.59 (1H, d, J=9 Hz), 12.64 (1H, broad s).

(3) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 1.35 g.), conc. hydrochloric acid (3.926 g.), methanol(20 ml.), water (10 ml.) and tetrahydrofuran (40 ml.) was stirred at 30°C. for 6 hours. The resultant solution was concentrated in vacuo inorder to evaporate the methanol, and the aqueous solution obtained wasadjusted to pH 4.2 with 10% aqueous solution of sodium hydroxide. Thesolution was adjusted to pH 3.0 with 10% hydrochloric acid. Theprecipitates were collected by filtration and dried to give7-[2-(2-aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 0.8 g.).

I.R. ν_(max) ^(Nujol) : 3300 (broad), 3200 (broad), 1775, 1670, 1635cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.64 (2H, s), 4.64 (2H, s), 5.13 (1H, d, J=5Hz), 5.86 (1H, dd, J=5 Hz, 7 Hz), 6.49 (1H, t, J=4 Hz), 6.82 (1H, s),7.33 (2H, s), 9.57 (1H, d, J=9 Hz).

EXAMPLE 33

(1) A solution of2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetic acid(syn isomer, 0.9 g.), N,N-dimethylformamide (0.24 g.) and phosphorylchloride (0.5 g.) in ethyl acetate (10 ml.) and a solution of4-nitrobenzyl 7-amino-3-chloro-3-cephem-4-carboxylate hydrochloride(1.23 g.) and trimethylsilylacetamide (2.8 g.) in ethyl acetate (20 ml.)were treated in a similar manner to that of Example 21-(1) to give4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 1.9 g.).

I.R. ν_(max) ^(Nujol) : 3250, 1790, 1740, 1700, 1660, 1610, 1530 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.92 (2H, q, J=17 Hz), 4.77 (2H, q, J=8.5 Hz),5.35 (1H, d, J=5 Hz), 5.48 (2H, s), 5.95 (1H, dd, J=5 Hz, 8 Hz), 7.50(1H, s), 7.70 (2H, d, J=9 Hz), 8.27 (2H, d, J=9 Hz), 8.53 (1H, s), 9.92(1H, d, J=8 Hz), 12.67 (1H, broad s).

(2) A suspension of 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 1.8 g.) and 10% palladium carbon (0.9 g.) in methanol (20ml.) and tetrahydrofuran (20 ml.) was treated in a similar manner tothat of Example 21-(2) to give7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.0 g.).

I.R. ν_(max) ^(Nujol) : 3250, 1780, 1690, 1655, 1530 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.86 (2H, q, J=17 Hz), 4.80 (2H, q, J=8.5 Hz),5.33 (1H, d, J=5 Hz), 5.92 (1H, dd, J=5 Hz, 8 Hz), 7.53 (1H, s), 8.53(1H, s), 9.93 (1H, d, J=8 Hz), 12.70 (1H, broad s).

(3)7-[2-(2-Formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 0.7 g.), conc.hydrochloric acid (0.43 ml.) andmethanol (16 ml.) were treated in a similar manner to that of Example21-(3) to give7-[2-(2-aminothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 0.65 g.)

I.R. ν_(max) ^(Nujol) : 3320, 3150, 1775, 1720, 1660, 1645, 1600, 1545cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.87 (2H, q, J=18 Hz), 4.80 (2H, q, J=8.5 Hz),5.30 (1H, d, J=5 Hz), 5.83 (1H, dd, J=5 Hz, 8 Hz), 7.05 (1H, s), 10.00(1H, d, J=8 Hz).

EXAMPLE 34

(1) A solution of 2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetic acid(syn isomer, 1.5 g.), N,N-dimethylformamide (440 mg.) and phosphorylchloride (920 mg.) in ethyl acetate (12 ml.) and a solution of4-nitrobenzyl 7-amino-3-chloro-3-cephem-4-carboxylate (2.03 g.),trimethylsilylacetamide (7 g.) and bis(trimethylsilyl)acetamide (2 ml.)in ethyl acetate (25 ml.) were treated in a similar manner to that ofExample 21-(1) to give 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 2.8 g.), yellow powder.

I.R. ν_(max) ^(Nujol) : 3200-3250, 1780, 1730, 1690, 1655, 1605, 1530,1350 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.90 (3H, m), 1.2-1.6 (4H, m), 3.88 (2H, AB-q,J=19 Hz), 4.0-4.2 (2H, m), 5.32 (1H, d, J=4 Hz), 5.44 (2H, s), 5.92 (1H,d,d, J=4 Hz, 8 Hz), 7.36 (1H, s), 7.68 (2H, d, J=8 Hz), 8.22 (2H, d, J=8Hz), 8.50 (1H, s), 9.72 (1H, d, J=8 Hz), 12.56 (1H, s).

(2) A mixture of 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 2.7 g.), 10% palladium carbon (1.3 g.), water (4 ml.),acetic acid (0.4 ml.), methanol (27 ml.) and tetrahydrofuran (54 ml.)was treated in a similar manner to that of Example 21-(2) to give7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.4 g.), pale yellow powder.

I.R. ν_(max) ^(Nujol) : 3250, 2400-2600, 1780, 1700, 1690, 1650, 1610cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.90 (3H, m), 1.2-1.70 (4H, m), 3.88 (2H, ABq,J=19 Hz), 4.0-4.25 (2H, m), 5.32 (1H, d, J=5 Hz), 5.90 (1H,d,d, J=5 Hz,9 Hz), 7.42 (1H, s), 8.50 (1H, s), 9.73 (1H, d, J=8 Hz), 12.60 (1H, s).

(3) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.3 g.), conc. hydrochloric acid (1.3 ml.) andmethanol (20 ml.) was treated in a similar manner to that of Example21-(3) to give7-[2-(2-aminothiazol-4-yl)-2-butoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.2 g.), pale yellow powder.

I.R. ν_(max) ^(Nujol) : 3300, 2500-2600, 1785, 1730, 1655, 1630 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.90 (3H, m), 1.2-1.75 (4H, m), 3.88 (2H, AB-q,J=19 Hz), 5.17 (2H, m), 5.33 (1H, d, J=5 Hz), 5.83 (1H, d,d, J=5 Hz, 8Hz), 6.93 (1H, s), 9.50 (2H, broad s), 9.85 (1H, d, J=8 Hz).

EXAMPLE 35

(1) A solution of 2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetic acid(syn isomer, 1.09 g.), N,N-dimethylformamide (322 mg.) and phosphorylchloride (675 mg.) in ethyl acetate (9.2 ml.) and a solution of4-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate hydrochloride(1.5 g.), trimethylsilylacetamide (5 g.) andbis(trimethylsilyl)acetamide (2 ml.) in ethyl acetate (30 ml.) weretreated in a similar manner to that of Example 3-(1) to give4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylate(syn isomer,1.8 g.).

I.R. ν_(max) ^(Nujol) : 3300, 3220, 1770, 1715, 1690, 1650, 1610, 1540,1350 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.90 (3H, m), 1.2-1.7 (4H, m), 3.72 (2H, broads), 3.96 (3H, s), 4.10 (2H, m), 5.22 (1H, d, J=4 Hz), 5.32 (2H, s), 5.75(1H, d,d, J=4 Hz, 8 Hz), 7.43 (1H, s), 7.64 (2H, d, J=9 Hz), 8.20 (2H,d, J=9 Hz), 8.48 (1H, s), 9.56 (1H, d, J=8 Hz), 12.59 (1H, s).

(2) A mixture of 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylate(syn isomer, 1.7 g.), 10% palladium carbon (1 g.), water (3 ml.), aceticacid (0.3 ml.), methanol (20 ml.) and tetrahydrofuran (35 ml.) wastreated in a similar manner to that of Example 15-(2) to give7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer, 1 g.), yellow powder.

I.R. ν_(max) ^(Nujol) : 3200-3250, 2600, 1775, 1700, 1690, 1650 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 1.0 (3H, m), 1.2-1.75 (4H, m), 3.67 (2H, broads), 3.86 (3H, s), 4.0-4.3 (2H, m), 5.23 (1H, d, J=4 Hz), 5.68 (1H, d,d,J=4 Hz, 8 Hz), 7.50 (1H, s), 8.58 (1H, s), 9.63 (1H, d, J=8 Hz).

(3) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer, 0.9 g.), conc. hydrochloric acid (0.8 ml.) andmethanol (13.5 ml.) was treated in a similar manner to that of Example15-(3) to give7-[2-(2-aminothiazol-4-yl)-2-butoxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer, 0.4 g.), yellowish white powder.

I.R. ν_(max) ^(Nujol) : 3200-3300, 2600, 1770, 1705, 1670, 1620 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.90 (3H, m), 1.2-1.65 (4H, m), 3.60 (2H, s),3.96 (3H, s), 4.0-4.16 (2H, m), 5.12 (1H, d, J=4 Hz), 5.55 (1H, d,d J=4Hz, 8 Hz), 6.80 (1H, s), 7.2-7.6 (2H, broad s), 9.50 (1H, d, J=8 Hz).

EXAMPLE 36

(1) 4-Nitrobenzyl 7-amino-3-cephem-4-carboxylate (5 g.) was dissolved ina solution of trimethylsilylacetamide (13.8 g.) andbis(trimethylsilyl)acetamide (10 ml.) in dry ethyl acetate (50 ml.) andstirred at 45° C. for 1.5 hours. A solution of bromine (2.88 g.) inmethylene chloride (7 ml.) was added dropwise to a solution of diketene(1.5 g.) in methylene chloride (7 ml.) at -40° C. over 20 minutes andstirred at -30° C. for 1 hour. The solution obtained thus was added todropwise to the above solution of 4-nitrobenzyl7-amino-3-cephem-4-carboxylate under cooling at -15° C. and then stirredat the same temperature for 30 minutes. Water (50 ml.) was added to theresultant solution and extracted with ethyl acetate. The ethyl acetateextract was washed with water, dried over magnesium sulfate andconcentrated under reduced pressure to give oily 4-nitrobenzyl7-[2-(2-bromoacetyl)acetamido]-3-cephem-4-carboxylate (6.15 g.).

I.R. ν_(max) ^(Nujol) : 1780, 1740, 1630 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.62 (2H, broad s), 4.37 (2H, s), 5.08 (1H, d,J=5 Hz), 5.40 (2H, s), 5.77-6.05 (m), 6.67 (1H, t, J=5 Hz), 7.68, 8.04(4H, m, J=9 Hz), 9.07 (1H, d, J=8 Hz).

(2) 4-Nitrobenzyl 7-[2-(2-bromoacetyl)acetamido]-3-cephem-4-carboxylate(8.40 g.) was suspended in a mixture of tetrahydrofuran (150 ml.) andwater (30 ml.) To the suspension were added acetic acid (50 ml.) and asolution of sodium nitrite (1.20 g.) in water (15 ml.) underice-cooling, and stirred at 20° to 22° C. for 1.5 hours. The resultantsolution was poured into ice-water (300 ml.) and stirred for 20 minutes.The precipitating substance was collected by filtration, washed withwater, dried and then recrystallized from ethyl acetate to give4-nitrobenzyl7-[2-(2-bromoacetyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 3.1 g.), mp 153° to 162° C.

I.R. ν_(max) ^(Nujol) : 3250, 1780, 1720, 1705, 1650, 1610, 1600(shoulder), 1550, 1520 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.67 (2H, d, J=4 Hz), 4.63 (1.5H, s), 4.88 (0.5H,s), 5.18 (1H, d, J=5 Hz), 5.45 (2H, s), 5.93 (1H, dd, J=5 Hz, 8 Hz),6.72 (1H, t, J=4 Hz), 7.73 (2H, d, J=9 Hz), 8.28 (2H, d, J=9 Hz), 9.38(1H, d, J=8 Hz), 11.27 (1H, s).

(3) A solution of diazomethane in diethyl ether was added little bylittle to a solution of 4-nitrobenzyl7-[2-(2-bromoacetyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylate(0.9 g.) in tetrahydrofuran (30 ml.) under ice-cooling until thereaction terminated, and then acetic acid was added to the resultantsolution to decompose excess diazomethane. The resultant solution wasconcentrated under reduced pressure to give the foamy product of4-nitrobenzyl7-[2-(2-bromoacetyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 0.9 g.)

(4) Thiourea (0.14 g.) was added to a solution of 4-nitrobenzyl7-[2-(2-bromoacetyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 0.8 g.) in ethanol (20 ml.) and water (5 ml.), and stirredat room temperature for 3.5 hours. The resultant solution wasconcentrated under reduced pressure, and to the residue were added waterand ethyl acetate. The ethyl acetate layer was separated, washed withwater, dried over magnesium sulfate and concentrated under reducedpressure to give the crude product (0.6 g.) The product was purified bycolumn chromatography on silica gel (eluent: benzene and ethyl acetate(8:2) to give 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 0.21 g.), mp 165° to 170° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3350-3200, 1770, 1720, 1665, 1615, 1515 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.60 (2H, broad s), 3.81 (3H, s), 5.12 (1H, d,J=5 Hz), 5.36 (2H, s), 5.83 (1H, dd, J=5 Hz, 8 Hz), 6.64 (1H, t, J=4Hz), 6.70 (1H, s), 7.20 (2H, s), 7.65 (2H, d, J=9 Hz), 8.19 (2H, d, J=9Hz), 9.60 (1H, d, J=8 Hz).

EXAMPLE 37

(1) A solution of bromine (43.0 g.) in methylene chloride (30 ml.) wasdropwise added to a solution of diketene (22.6 g.) in methylene chloride(30 ml.) at -30° C. over 35 minutes, and stirred at the same temperaturefor 30 minutes. The solution was dropwise added to a stirred solution of4-nitrobenzyl 7-amino-3-cephem-4-carboxylate (75.1 g.) andbis(trimethylsilyl)acetamide (68.4 g.) in tetrahydrofuran (1.5 l) at-15° C. over 10 minutes, and the solution was stirred at the sametemperature for 50 minutes. Water (35 ml.) and an aqueous solution (35ml.) of sodium nitrite (18.6 g.) were added to the resultant solutionwhile keeping at pH 2.0, and the solution was stirred at 10° to 15° C.for 15 minutes. After the solution was adjusted to pH 4.5 with asaturated aqueous solution of sodium bicarbonate, an aqueous solution(150 ml.) of thiourea (17.1 g.) was added to the solution, adjusted topH 6.0 with a saturated aqueous solution of sodium bicarbonate, andstirred for 20 minutes. The organic layer was separated and concentratedunder reduced pressure. The residue was dissolved in ethyl acetate (1.5l), and washed with water three times. The solution was dried overmagnesium sulfate, treated with activated charcoal and concentratedunder reduced pressure. After the residue was triturated with diethylether (200 ml.), the precipitates were collected by decantation andwashed with ether acetate (300 ml.), a mixture of tetrahydrofuran (500ml.) and ethyl acetate (1 l) at 60° C. and then with ethyl acetate (100ml.) three times, and dried to give 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 55.5 g.).

I.R. ν_(max) ^(Nujol) : 1760, 1710, 1660, 1630 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.60 (2H, d, J=5 Hz), 5.12 (1H, d, J=5 Hz),5.39(2H, s), 5.88 (1H, dd, J=8 Hz, 5 Hz), 6.63 (1H, s), 6.53-6.77 (1H,m), 7.08 (2H, broad s), 7.68 (2H, d, J=9 Hz), 8.22 (2H, d, J=9 Hz), 9.47(1H, d, J=8 Hz), 11.33 (1H, s).

(2) 10% Palladium carbon (0.35 g.) was added to a solution of4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 0.7 g.) in methanol (70 ml.), and the mixture was subjectedto catalytic reduction at room temperature under atmospheric pressurefor 1.5 hours. The resultant mixture was filtered, and the filtrate wasconcentrated under reduced pressure. To the residue was added an aqueoussolution of sodium bicarbonate and the insoluble substance was filteredout. The filtrate was washed with ethyl acetate and methylene chloridein turn, bubbled with nitrogen gas and then lyophilized. The residue wasdissolved in water (30 ml.) and adjusted to pH 3.8 with 10% hydrochloricacid. The solution was subjected to column chromatography onmacroporous, nonionic adsorption resin "Diaion HP-20" (Trade mark;manufactured by Mitsubishi Chemical Industries Ltd., 20 ml.), washedwith water, and then eluted with 40% aqueous acetone. After acetone wasremoved from the eluate under reduced pressure, the residue waslyophilized to give7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylicacid (syn-isomer: 0.25 g.)

I.R. ν_(max) ^(Nujol) : 3350 to 3200, 1770, 1670, 1630 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.60 (2H, broad s), 5.10 (1H, d, J=5 Hz), 5.83(1H, dd, J=5 Hz, 8 Hz), 6.47 (1H, t, J=4 Hz), 6.67 (1H, s), 9.47 (1H, d,J=8 Hz).

EXAMPLE 38

(1) Thiourea (0.18 g.) was added to a suspension of 4-nitrobenzyl7-[2-(2-bromoacetyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 1.05 g.) in ethanol (25 ml.), tetrahydrofuran (25 ml.) andwater (5 ml.), and stirred at room temperature for 4 hours. Theresultant solution was concentrated under reduced pressure and cooled.The residue was crystallized by treating with a mixture oftetrahydrofuran and ethyl acetate, and collected by filtration to give4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 0.95 g.), colorless crystals, mp 172° to 175° C. (dec.)

I.R. ν_(max) ^(Nujol) : 3350-3200, 1770, 1725, 1670, 1625, 1520 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.68 (2H, d, J=4 Hz), 5.20 (1H, d, J=5 Hz), 5.43(2H, s), 5.90 (1H, dd, J=8 Hz, 5 Hz), 6.70 (1H, t, J=4 Hz), 6.88 (1H,s), 7.70 (2H, d, J=9 Hz), 8.23 (2H, d, J=9 Hz), 9.68 (1H, d, J=8 Hz).

(2) A solution of diazomethane in diethyl ether was added little bylittle to a solution of 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-hydroxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 0.3 g.) in methanol (30 ml.) until the reaction terminated.The resultant solution was concentrated under reduced pressure, and theresidue was pulverized with diethyl ether, collected by filtration anddried to give 4-nitrobenzyl7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn-isomer: 0.26 g.). This product was identified with the authenticsample.

EXAMPLE 39

7-Amino-3-cephem-4-carboxylic acid (2.54 g.) was dissolved in a solutionof trimethylsilylacetamide (11.7 g.) and bis(trimethylsilyl)acetamide(15 ml.) in dried ethyl acetate (50 ml.) A solution of bromine (2.43 g.)in dried methylene chloride (10 ml.) was added dropwise to a solution ofdiketene (1.28 g.) in dried methylene chloride (25 ml.) at -30° C. over10 minutes and stirred at the same temperature for 1.5 hours. Thesolution was added to the above solution containing7-amino-3-cephem-4-carboxylic acid at -15° C. over 10 minutes, andstirred at -15° to -10° C. for 1.5 hours. Water (50 ml.) was added tothe resultant solution. The ethyl acetate layer was separated, andextracted with aqueous solution of sodium bicarbonate. The aqueousextract was adjusted to pH 2.0 with 10% hydrochloric acid and extractedwith ethyl acetate. The ethyl acetate extract was washed with water,dried over magnesium sulfate and concentrated under reduced pressure togive 7-[2-(2-bromoacetyl)acetamido]-3-cephem-4-carboxylic acid (2.82 g.)

I.R. ν_(max) ^(Nujol) : 1760, 1660 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.58 (2H, d, J=4 Hz), 3.65 (2H, s), 4.40 (2H, s),5.06 (1H, d, J=5 Hz), 5.73 (1H, dd, J=8 Hz, 5 Hz), 6.50 (1H, t, J=4 Hz),9.08 (1H, d, J=8 Hz).

EXAMPLE 40

The following compounds were prepared in a similar manner to that ofExample 36.

(1) 4-Nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-ethoxyiminoacetamido]-3-cephem-4carboxylate(syn isomer)

(2) 4-Nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-propoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

(3) 4-Nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-isopropoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

EXAMPLE 41

The following compounds were prepared in a similar manner to that ofExample 38-(2).

(1) 4-Nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-ethoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

(2) 4-Nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-propoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

(3) 4-Nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-isopropoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer)

EXAMPLE 42

(1) Sodium boron hydride (160 mg.) was added to a suspension of4-nitrobenzyl7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer, 1 g.) in tetrahydrofuran (10 ml.), acetic acid (3 ml.) andwater (1 ml.) at 0° C. over 10 minutes, and stirred at 0° to 3° C. for55 minutes. After water was added to the resultant solution, thesolution was extracted with ethyl acetate. The extract was washed with asaturated aqueous solution of sodium chloride, a saturated aqueoussolution of sodium bicarbonate and a saturated aqueous solution ofsodium chloride in turn, and dried over magnesium sulfate. The solutionwas concentrated under reduced pressure, and the residue was pulverizedwith diethyl ether to give 4-nitrobenzyl7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-hydroxycepham-4-carboxylate(syn isomer, 0.77 g.), mp 172° to 175° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3250, 1775, 1745, 1660 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 2.76 (1H, dd, J=14 Hz, 3 Hz), 3.17 (1H, dd, J=14Hz, 13 Hz), 3.92 (3H, s), 4.03 (1H, m), 4.72 (1H, d, J=6 Hz), 5.24 (1H,d, J=4 Hz), 5.37 (2H, s), 5.56 (1H, dd, J=9 Hz, 4 Hz), 6.07 (1H, d, J=4Hz), 7.44 (1H, s), 7.72 (2H, d, J=8 Hz), 8.27 (2H, d, J=8 Hz), 8.54 (1H,s), 9.67 (1H, d, J=9 Hz).

(2) Mesyl chloride (0.406 g.) was dropwise added to a stirred mixture of4-nitrobenzyl7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-hydroxycephem-4-carboxylate(syn isomer, 1 g.), N,N-dimethylformamide (10 ml.) and potassiumcarbonate (0.732 g.) at 0° to 5° C. over 2 minutes, and the solution wasstirred at room temperature for 2.5 hours. After ethyl acetate and waterwere added to the resultant solution, the solution was extracted withethyl acetate. The remaining aqueous layer was extracted again withethyl acetate. The ethyl acetate extract solution was washed with asaturated aqueous solution of sodium chloride, dried over magnesiumsulfate, and concentrated under reduced pressure. The residue wassubjected to column chromatography on silica gel (30 g.) and eluted witha mixture of chloroform and ethyl acetate. The eluate was concentratedunder reduced pressure to give 4-nitrobenzyl7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer, 0.12 g.), mp 224° C. (dec.).

EXAMPLE 43

Phosphoryl chloride (704 mg.) was added dropwise to a solution ofN,N-dimethylformamide (336 mg.) in ethyl acetate (8 ml.) below 5° C. andstirred at the same temperature for 30 minutes.2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetic acid (syn isomer, 1 g.)was added to the solution and stirred at 5° to 10° C. for an hour. Thesolution was added dropwise to a solution of7-amino-3-hydroxycepham-4-carboxylic acid (872 mg.) andtrimethylsilylacetamide (1.05 g.) in ethyl acetate (20 ml.) at -20° C.over 5 minutes, and stirred at -20° to -25° C. for an hour. Water (50ml.) was added to the resultant solution and adjusted to pH 7.0 withsodium bicarbonate. The aqueous layer was separated, and the ethylacetate layer was extracted again with water (10 ml.). The aqueousextracts were combined, adjusted to pH 6 and absorbed on macroporousnonionic adsorption resin Diaion HP-20 (50 ml., Trademark, manufacturedby Mitsubishi Chemical Industries). The column was washed with water (50ml.) and eluted with 30% aqueous isopropyl alcohol. The eluatecontaining the object compound was adjusted to pH 6.5 and concentratedunder reduced pressure. The residue was lyophilized to give sodium7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-hydroxycepham-4-carboxylate(syn isomer, 1.1 g.)

N.M.R. δ(D₂ O, ppm): 2.72-3.18 (2H, m), 4.02 (3H, s), 4.02-4.28 (1H, m),4.54 (1H, d, J=4 Hz), 5.28 (1H, d, J=4 Hz), 5.53 (1H, d, J=4 Hz), 7.50(1H, s), 8.53 (1H, s).

EXAMPLE 44

Thionyl chloride (0.423 g.) was dropwise added to a stirred solution of4-nitrobenzyl7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-hydroxy-3-cephem-4-carboxylate(syn isomer, 1 g.) in N,N-dimethylformamide (10 ml.) under ice coolingover 2 minutes, and the solution was stirred at room temperature for 1.1hours. Ethyl acetate (40 ml.) and water (30 ml.) were added to theresultant solution and shaken sufficiently. The aqueous layer wasextracted with ethyl acetate, and the extract was combined with theethyl acetate layer separated above. The ethyl acetate solution waswashed with a saturated aqueous solution of sodium chloride, dried overmagnesium sulfate and then concentrated under reduced pressure. Theresidue was subjected to column chromatography on silica gel (30 g.) andeluted with chloroform and then a mixture of chloroform and ethylacetate (7:3). The latter eluate was concentrated under reduced pressureto give 4-nitrobenzyl7-[2-(2-formamido-4-thiazolyl)-2-methoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 0.2 g.), mp 216° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3230 (shoulder), 3110, 3050, 1785, 1725, 1690,1655 cm⁻¹.

N.M.R. δppm (DMSO-d₆): 3.93 (3H, s), 3.93 (2H, q, J=18 Hz), 5.36 (1H, d,J=5 Hz), 5.50 (2H, s), 5.97 (1H, dd, J=5 Hz, 9 Hz), 7.45 (1H, z), 7.73(2H, d, J=9 Hz), 8.29 (2H, d, J=9 Hz), 8.56 (1H, s), 9.78 (1H, d, J=9Hz).

EXAMPLE 45

The following compounds were prepared in a similar manner to that ofExample 44.

(1) 4-Nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-propoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer)

(2) 4-Nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer)

(3) 4-Nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-butoxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer)

EXAMPLE 46 Fermentation

Pre-culture medium: Trypticase soy broth (BBL)

Main culture medium:

glycerin: 3 g

peptone: 1 g

corn steep liquor: 1 g

dry yeast: 2 g

sodium carbonate: 0.1 g

KH₂ PO₄ : 0.55 g

Na₂ HPO₄.12H₂ O: 2.15 g

(The above components were dissolved in water so as to become 100 ml intotal, and the medium was adjusted to pH 7.2.)

A main culture broth (100 ml.) was placed in Sakaguchi-flask (500 ml)and sterilized at 120° C. for 20 minutes. Into this medium, there wasinoculated a culture broth (1 ml.) of each of microorganisms as givenbelow, which were cultured in a pre-culture medium, respectively.

Reaction

Into above cultured broth, there was added the Substrate (0.1 g) asgiven below suspended in 0.1M phosphate buffer (pH 7.2, 1 ml.), and thenthe mixture was shaked at 30° C. for 48 hours.

Identification and assay

After the reaction, in order to identify the generated product thereaction mixture as obtained above was chromatographed on Eastmanchromatogram 6065 cellulose at room temperature. As a developing agent,there was used (A) the upper layer of a mixture of n-butanol, ethanoland water (4:1:5 by volume) and (B) a mixture of n-propanol and water(7:3 by volume). Rf value was determined by index of antimicrobialactivity against a sensitive strain of Escherichia coli ES 111, and asthe result only one spot showing each of Product I and II was observedon the Eastman chromatogram 6065 cellulose without showing any spot ofeach of Substrate I and II. Rf value are shown in the following table.

    ______________________________________                                                         Developing Solvent                                                            A    B                                                       ______________________________________                                        Reaction Mixture   0.85   0.90                                                (Product I)                                                                   Reference          0.39   0.60                                                (Substrate I)                                                                 Reaction Mixture   0.90   0.92                                                (Product II)                                                                  Reference          0.36   0.54                                                (Substrate II)                                                                ______________________________________                                         Note:                                                                         Substrate I: 4nitrobenzyl                                                     7[2formamido-4-thiazolyl)-2-methoxyiminoacetamido3-cephem-4-carboxylate       (syn isomer)                                                                  Product I:                                                                    7[2(2-formamido-4-thiazolyl)-2-methoxyimino-acetamido3-cephem-4-carboxyli     acid (syn isomer)                                                             Substrate II: 4nitrobenzyl                                                    7[2amino-4-thiazolyl)-2-methoxyiminoacetamido3-cephem-4-carboxylate (syn      isomer)                                                                       Product II:                                                                   7[2(2-amino-4-thiazolyl)-2-methoxyimino-acetamido3-cephem-4-carboxylic        acid (syn isomer)                                                        

The product generated in the reaction mixture as obtained above wasassayed by paper disk-plate method using a sensitive strain ofEscherichia coli ES 111 (culture: 37° C., 16 hours) and the yieldthereof was calculated. The results are shown in the following.

    ______________________________________                                        Microorganism                                                                 used for enzymatic   Yield (%)                                                hydrolysis           Product I Product II                                     ______________________________________                                        Bacillus subtilis IAM 1069                                                                         75        60                                             Bacillus sphaericus IAM 1286                                                                       75        20                                             Bacillus subtilis IAM 1107                                                                         75        95                                             Bacillus subtilis IAM 1214                                                                         85        20                                             Corynebacterium equi IAM 1038                                                                      95        95                                             Micrococcus varians IAM 1314                                                                       70        20                                             Flavobacterium rigens IAM 1238                                                                     85        90                                             Salmonella typhimurium IAM 1406                                                                    90        20                                             Staphylococcus epidermidis IAM 1296                                                                90        95                                             Microbacterium flavum IAM 1642                                                                     90        95                                             ______________________________________                                    

The following are examples of pharmaceutical compositions prepared inaccordance with this invention and containing7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, Compound A) as the active substance.

EXAMPLE 47 (lyophilized preparation for injection)

Sodium salt of compound A (20 g. potency) was dissolved in water (200ml.), and the solution (5 ml.) was filled in each 10 ml. vial. Thesevials are frozen and dried in a vacuum (lyophilization).

EXAMPLE 48 (suspension for injection)

Compound A: 25 g.

Methyl cellurose: 0.5 g.

Methyl 4-oxobenzoate: 0.1 g.

Polysolvate 80: 0.1 g.

Lidocaine hydrochloride: 0.5 g.

Water for injection to make 100 ml.

This aqueous suspension is suitable for intramuscular injection.

EXAMPLE 49 (tablets for oral use)

Compound A: 500 mg.

Lactose: 375.5 mg.

Hydroxypropylcellurose: 2 mg.

Magnesium stearate: 22.5 mg.

This mixture provides a tablet for oral use in the treatment ofinfectious deseases caused by pathogenic bacteria.

EXAMPLE 50 (capsule for oral use)

Compound A: 500 mg.

Magnesium stearate: 10 mg.

This mixture provides a capsule for oral use in the treatment ofinfectious deseases caused by pathogenic bacteria.

EXAMPLE 51

(1) A solution of 2-(2-formamidothiazol-4-yl)-2-n-hexyloxyiminoaceticacid (syn isomer, 3.24 g.), N,N-dimethylformamide (0.951 g.), andphosphoryl chloride (2.00 g.) in ethyl acetate (20 ml.), and a solutionof 4-nitrobenzyl 7-amino-3-chloro-3-cephem-4-carboxylate hydrochloride(4 g.) in a mixture of acetone (20 ml.), water (20 ml.) andtetrahydrofuran (40 ml.) were treated in a similar manner to that ofExample 21-(1) to give 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 5.78 g.).

I.R. ν_(max) ^(Nujol) : 3240, 3200, 3050, 1780, 1730, 1695, 1660 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6˜2.1 (11H, m), 3.96 (2H, q, J=19 Hz), 4.15(2H, t, J=6 Hz), 5.37 (1H, d, J=5 Hz), 5.50 (2H, s), 5.97 (1H, d, d, J=5Hz, 8 Hz), 7.42 (1H, s), 7.72 (2H, d, J=9 Hz), 8.28 (2H, d, J=9 Hz),8.54 (1H, s), 9.74 (1H, d, J=8 Hz), 12.73 (1H, broad s).

(2) A mixture of 4-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 5.6 g.), acetic acid (0.4 ml.), 10% palladium carbon (2.24g.), water (4 ml.), methanol (23 ml.) and tetrahydrofuran (56 ml.) wastreated in a similar manner to that of Example 21-(2) to give7-[2-(2-formamidothiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 2.5 g.).

I.R. ν_(max) ^(Nujol) : 3255, 1785, 1690, 1650 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6˜2.0 (11H, m), 3.86 (2H, q, J=18 Hz), 4.13(2H, t, J=6 Hz), 5.30 (1H, d, J=5 Hz), 5.88 (1H, d, d, J=5 Hz, 8 Hz),7.41 (1H, s), 8.54 (1H, s), 9.70 (1H, d, J=8 Hz), 12.68 (1H, s).

(3) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 2.4 g.), conc.hydrochloric acid (1.84 g.), methanol(36 ml.) and tetrahydrofuran (30 ml.) was stirred at 30° C. for 2 hours.The resultant solution was concentrated in vacuo. Water (60 ml.) wasadded to the residue and the precipitates were collected by filtration,washed with water and dried over phosphorus pentoxide to give7-[2-(2-aminothiazol-4-yl)-2-n-hexyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.86 g.).

I.R. ν_(max) ^(Nujol) : 3300, 1780, 1665, 1535 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6˜2.0 (11H, m), 3.84 (2H, q, J=18 Hz), 4.08(2H, t, J=7 Hz), 5.28 (1H, d, J=5 Hz), 5.82 (1H, d, d, J=5 Hz, 8 Hz),6.77 (1H, s), 9.66 (1H, d, J=8 Hz), 6.0˜8.0 (2H, broad s).

EXAMPLE 52

(1) 2-(2-Formamidothiazol-4-yl)-2-propargyloxyiminoacetic acid (synisomer, 1.27 g.), N,N-dimethylformamide (402 mg.), phosphoryl chloride(843 mg.) and ethyl acetate (11.2 ml.) were treated in a conventionalmanner to prepare an activated acid solution. On the other hand, amixture of 7-amino-3-methoxy-3-cephem-4-carboxylic acid hydrochloride(1.33 g.), trimethylsilylacetamide (4 g.), bis(trimethylsilyl)acetamide(2 ml.) and ethyl acetate (20 ml.) was stirred at 40° to 45° C. for anhour. To the solution was added the activated acid solution obtainedabove all at once at -15° C., and then stirred at -5° to -10° C. for 1.5hours. Water (30 ml.) was added to the resultant solution, filtered andthe organic layer was separated. The insoluble substance filtered outwas dissolved in a saturated aqueous solution of sodium bicarbonate, andthe solution was added to the organic layer. The solution was adjustedto pH 7.5 and the aqueous solution was separated, and then extractedwith ethyl acetate at pH 2.0.

The extract was washed with water, dried over magnesium sulfate andfiltered. The filtrate was concentrated in vacuo to give7-[2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer, 1.0 g.), yellow powder.

I.R. ν_(max) ^(Nujol) : 3200˜3300, 2500˜2600, 2120, 1770, 1710, 1690,1670 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.50 (1H, m), 3.65 (2H, s), 3.82 (3H, s), 4.80(2H, d, J=2 Hz), 5.20 (1H, d, J=4 Hz), 5.62 (1H, d,d, J=4 Hz, 8 Hz),7.52 (1H, s), 8.55 (1H, s), 9.68 (1H, d, J=8 Hz), 12.65 (1H, broad s).

(2) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer, 0.9 g.), conc. hydrochloric acid (0.9 ml.) andmethanol (13.5 ml.) was stirred at room temperature for 4 hours. Afterconcentrating the resultant solution in vacuo at 35° C., the residue wasdissolved in water and washed with ethyl acetate. The aqueous solutionwas adjusted to pH 7.0 with sodium bicarbonate and washed with ethylacetate and diethyl ether. After removing the organic solvent bybubbling nitrogen gas, the solution was adjusted to pH 3.0 with 10%hydrochloric acid and stirred under ice cooling. The precipitates werecollected by filtration, washed with water and dried to give7-[2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-methoxy-3-cephem-4-carboxylicacid (syn isomer, 0.25 g.), whitish yellow powder.

I.R. ν_(max) ^(Nujol) : 3300, 2500˜2600, 2120, 1775, 1710, 1670, 1620cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.52 (1H, m), 3.82 (3H, s), 4.77 (2H, d, J=2Hz), 5.17 (1H, d, J=4 Hz), 5.58 (1H, d,d, J=4 Hz, 8 Hz), 6.93 (1H, s),7.1˜7.3 (2H, broad s), 9.67 (1H, d, J=8 Hz).

EXAMPLE 53

(1) 2-(2-Formamidothiazol-4-yl)-2-propargyloxyiminoacetic acid (synisomer, 1.27 g.), N,N-dimethylformamide (400 mg.), phosphoryl chloride(850 mg.) and ethyl acetate (11.2 ml.) were treated in a conventionalmanner to give the activated acid solution. The activated acid solutionwas added to a solution of 7-amino-3-chloro-3-cephem-4-carboxylic acidhydrochloride (2 g.), trimethylsilylacetamide (6 g.) andbis(trimethylsilyl)acetamide (3 ml.) in ethyl acetate (40 ml.) at -15°C. all at once, and stirred at -5° to -10° C. for 1.5 hours. Afteradding water (30 ml.) to the resultant solution, the ethyl acetate layerwas separated and extracted with a saturated aqueous solution of sodiumbicarbonate. Ethyl acetate was added to the aqueous extract, adjusted topH 2.0 with 10% hydrochloric acid. The ethyl acetate layer wasseparated, washed with water, dried over magnesium sulfate and filtered.The filtrate was concentrated in vacuo, and the residue was crystallizedwith a mixture of ethyl acetate and diisopropyl ether. The precipitateswere collected by filtration and dried to give7-[2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.5 g.), yellow powder.

I.R. ν_(max) ^(Nujol) : 3250˜3300, 2500˜2600, 2120, 1780, 1725, 1690,1670 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.45 (1H, m), 3.57 (2H, AB-q, J=20 Hz), 4.77(2H, d, J=2 Hz), 5.28 (1H, d, J=4 Hz), 5.80 (1H, d,d, J=4 Hz, 8 Hz),8.42 (1H, s), 8.52 (1H, s), 9.78 (1H, d, J=8 Hz), 12.72 (1H, broad s).

(2) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.4 g.), conc. hydrochloric acid (1.4 ml.) andmethanol (20 ml.) was treated in a similar manner to that of Example51-(2) to give7-[2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 0.7 g.), yellowish white powder.

I.R. ν_(max) ^(Nujol) : 3350, 2500˜2600, 2130, 1775, 1710, 1670, 1630cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 4.38 (1H, m), 4.48 (2H, AB-q, J=19 Hz), 4.72(2H, d, J=2 Hz), 5.28 (1H, d, J=4 Hz), 5.80 (1H, d,d, J=4 Hz, 8 Hz),6.78 (1H, s), 9.73 (1H, d, J=8 Hz).

EXAMPLE 54

Thiourea (11 mg.) and a solution of7-[2-(2-bromoacetyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylic acid(syn isomer, 30 mg.) in ethanol (2 ml.) were treated in a similar mannerto that of Example 36-(4) to give7-[2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer). The product was identified with an authentic sampleby thin layer chromatography.

EXAMPLE 55

(1) A solution of 4-nitrobenzyl 7-amino-3-chloro-3-cephem-4-carboxylatehydrochloride (15.0 g.), bis(trimethylsilyl)acetamide (11.3 g.) andtrimethylsilylacetamide (9.7 g.) in tetrahydrofuran (300 ml.), asolution of diketene (3.41 ml.) in methylene chloride (4 ml.), asolution of bromine (2.27 ml.) in methylene chloride (4 ml.), a solutionof sodium nitrite (3.1 g.) in water (20 ml.) and a solution of thiourea(4.0 g.) in water (40 ml.) were treated in a similar manner to that ofExample 37-(1) to give 4-nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-hydroxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 10.4 g.).

I.R. ν_(max) ^(Nujol) : 3300, 3180, 1777, 1730, 1670, 1603 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.93 (2H, d, J=5 Hz), 5.33 (1H, d, J=5 Hz), 5.49(2H, s), 5.90 (1H, d,d, J=5 Hz, 8.2 Hz), 6.68 (1H, s), 7.14 (1H, broads), 7.72 (2H, d, J=9.2 Hz), 8.27 (2H, d, J=9.2 Hz), 9.54 (1H, d, J=8.2Hz).

(2) 4-Nitrobenzyl7-[2-(2-aminothiazol-4-yl)-2-hydroxyiminoacetamido]-3-chloro-3-cephem-4-carboxylate(syn isomer, 5.0 g.), 10% palladium carbon (3.0 g.), methanol (100 ml.),water (10 ml.) and tetrahydrofuran (150 ml.) were treated in a similarmanner to that of Example 37-(2), to give7-[2-(2-aminothiazol-4-yl)-2-hydroxyiminoacetamido]-3-chloro-3-cephem-4-carboxylicacid (syn isomer, 1.28 g.).

I.R. ν_(max) ^(Nujol) : 3330, 1775, 1675, 1630 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 3.72 (2H, m), 5.24 (1H, d, J=5 Hz), 5.80 (1H,d,d, J=5.0 Hz, 8.2 Hz), 6.66 (1H, s), 9.50 (1H, d).

EXAMPLE R

(1) Ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 100 g.),N,N-dimethylformamide (300 ml.), potassium carbonate (130 g.) andbromooctane (121 g.) were treated in a similar manner to that of ExampleF-(1) to give ethyl 2-n-octyloxyimino-3-oxobutyrate (syn isomer, 165.5g.), oil.

I.R. ν_(max) ^(film) : 1745, 1695, 1470 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 0.6˜2.1 (18H, m), 2.35 (3H, s), 4.0˜4.6 (4H, m).

(2) Ethyl 2-n-octyloxyimino-3-oxobutyrate (syn isomer, 165.5 g.)sulfuryl chloride (84.7 g.) and acetic acid (165 ml.) were treated in asimilar manner to that of Example F-(2) to give ethyl2-n-octyloxyimino-4-chloro-3-oxobutyrate (syn isomer, 169.6 g.), oil.

I.R. ν_(max) ^(film) : 1745, 1710, 1465 cm⁻¹.

N.M.R. δ(CCl₄, ppm): 0.6˜2.1 (18H, m), 4.0˜4.6 (4H, m), 4.48 (2H, s).

(3) Ethyl 2-n-octyloxyimino-4-chloro-3-oxobutyrate (syn isomer, 169.6g.), thiourea (42.3 g.), sodium acetate trihydrate (75.5 g.), water (420ml.) and ethanol (1020 ml.) were treated in a similar manner to that ofExample F-(3) to give ethyl2-(2-aminothiazol-4-yl)-2-n-octyloxyiminoacetate (syn isomer, 65 g.),mp. 77° to 78° C.

I.R. ν_(max) ^(Nujol) : 3470, 3250, 3125, 1735, 1545, 1465 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.81 (3H, t, J=6 Hz), 0.6˜1.9 (15H, m), 4.07(2H, t, J=6 Hz), 4.28 (2H, q, J=7 Hz), 6.86 (1H, s), 7.02 (2H, broad s).

(4) Ethyl 2-(2-aminothiazol-4-yl)-2-n-octyloxyiminoacetate (syn isomer,64 g.), 2N-aqueous solution of sodium hydroxide (196 ml.), methanol (196ml.) and tetrahydrofuran (300 ml.) were treated in a similar manner tothat of Example F-(4) to give2-(2-aminothiazol-4-yl)-2-n-octyloxyiminoacetic acid (syn isomer, 52.5g.), mp. 146° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3170, 1635, 1565, 1460 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.86 (3H, t, J=6 Hz), 0.6˜1.9 (12H, m), 4.06(2H, t, J=6 Hz), 6.81 (1H, s), 7.22 (2H, s).

(5) 2-(2-Aminothiazol-4-yl)-2-n-octyloxyiminoacetic acid (syn isomer, 20g.), acetic anhydride (27.3 g.) and formic acid (12.3 g.) were treatedin a similar manner to that of Example F-(5) to give2-(2-formamidothiazol-4-yl)-2-n-octyloxyiminoacetic acid (syn isomer,21.3 g.), mp. 122° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3350, 3150, 3050, 1700, 1675, 1560 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6˜2.0 (15H, m), 4.16 (2H, t, J=6 Hz), 7.56(1H, s), 8.57 (1H, s), 12.67 (1H, s).

EXAMPLE 56

(1) A solution of 2-(2-formamidothiazol-4-yl)-2-n-octyloxyiminoaceticacid (syn isomer, 7.52 g.), phosphoryl chloride (5.4 g.) andN,N-dimethylformamide (2.58 g.) in tetrahydrofuran (16 ml.), which wasprepared in a similar manner to that of Example 30-(1), and a solutionof 7-amino-3-cephem-4-carboxylic acid (4 g.) in a mixture of acetone (20ml.) water (20 ml.) and tetrahydrofuran (20 ml.) were treated in asimilar manner to that of Example 30-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-n-octyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 8.1 g.).

I.R. ν_(max) ^(Nujol) : 3280, 3200, 3060, 1795, 1705, 1660, 1630 cm⁻¹.

N.M.R. δ(DMSO-d₆, ppm): 0.6˜2.1 (15H, m), 3.62 (2H, d, J=4 Hz), 4.14(2H, t, J=6 Hz), 5.16 (1H, d, J=5 Hz), 5.88 (1H, d,d, J=5 Hz, 8 Hz),6.51 (1H, t, J=4 Hz), 7.42 (1H, s), 8.54 (1H, s), 9.63 (1H, d, J=8 Hz),12.66 (1H, s).

(2)7-[2-(2-Formamidothiazol-4-yl)-4-n-octyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 8.0 g.), conc. hydrochloric acid (6.23 g.),tetrahydrofuran (15 ml.) and methanol (120 ml.) were treated in asimilar manner to that of Example 21-(3) to give7-[2-(2-aminothiazol-4-yl)-2-n-octyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer, 6.95 g.).

I.R. ν_(max) ^(Nujol) : 3320 (shoulder), 1785, 1660, 1630, 1535.

N.M.R. δ(DMSO-d₆, ppm): 0.6˜2.0 (15H, m), 3.62 (2H, broad s), 4.07 (2H,t, J=6 Hz), 5.12 (1H, d, J=5 Hz), 5.83 (1H, d,d, J=5 Hz, 9 Hz), 6.48(1H, broad s), 6.72 (1H, s), 7.22 (2H, s), 9.53 (1H, d, J=9 Hz).

What we claim is :
 1. A syn compound of the formula: ##STR37## whereinR² is an aliphatic hydrocarbon residue substituted with halogen, carboxyor esterified carboxy, or cycloalkyl;R⁵ is carboxy or functionallymodified carboxy and R⁶ is amino or protected amino, andpharmaceutically acceptable salt thereof.
 2. A syn compound of theformula: ##STR38## wherein R² is alkyl substituted with halogen, carboxyor esterified carboxy; or cyclo (lower) alkyl;R⁵ is carboxy orfunctionally modified carboxy and R⁶ is amino or protected amino, andpharmaceutically acceptable salt thereof.
 3. A syn compound of theformula: ##STR39## wherein R² is cycloalkyl,R⁵ is carboxy or esterifiedcarboxy and R⁶ is amino or protected amino, and pharmaceuticallyacceptable salt thereof.
 4. A syn compound of the formula: ##STR40##wherein R² is alkyl substituted with halogen, carboxy or loweralkoxycarbonyl,R⁵ is carboxy or esterified carboxy and R⁶ is amino orprotected amino, and pharmaceutically acceptable salt thereof.
 5. Thecompound of the claim 3, wherein R⁶ is amino or lower alkanamido.
 6. Thecompound of the claim 5, wherein R⁶ is amino or formamido and R⁵ iscarboxy.
 7. The compound of claim 6, which is7-[2-(2-aminothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 8. The compound of the claim 6, which is7-[2-(2-formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 9. The compounds of the claim 5, wherein R⁶ is aminoor formamido and R⁵ is substituted benzyloxycarbonyl.
 10. The compoundof the claim 9, which is p-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-cyclohexyloxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer).
 11. The compound of the claim 4, wherein R² is haloalkyl,and R⁶ is amino or lower alkanamido.
 12. The compound of the claim 11,wherein R² is monohalo(lower)alkyl or trihalo(lower)alkyl, and R⁵ iscarboxy.
 13. The compound of the claim 12, which is7-[2-(2-aminothiazol-4-yl)-2-(2-chloroethoxy)iminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 14. The compound of the claim 12, which is7-[2-(2-formamidothiazol-4-yl)-2-(2-chloroethoxy)iminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 15. The compound of the claim 12, which is7-[2-(2-aminothiazol-4-yl)-2-(2,2,2-trifluoroethoxy)iminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 16. The compound of the claim 12, which is7-[2-(2-formamidothiazol-4-yl)-2-(2,2,2-trifluoroethoxy)iminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 17. The compound of the claim 1, wherein R² iscarboxy(lower)alkyl and R⁶ is amino or lower alkanamido.
 18. Thecompound of the claim 17, which is7-[2-(2-aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 19. The compound of the claim 17, which is7-[2-(2-formamidothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 20. The compound of the claim 19, wherein R² is loweralkoxycarbonyl(lower)alkyl and R⁶ is amino or lower alkanamido.
 21. Thecompound of the claim 20, wherein R⁵ is carboxy.
 22. The compound of theclaim 21, which is7-[2-(2-aminothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 23. The compound of the claim 20, which is7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 24. The compound of the claim 20, which is7-[2-(2-formamidothiazol-4-yl)-2-tertbutoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylicacid (syn isomer).
 25. The compound of the claim 20, wherein R⁵ issubstituted benzyloxycarbonyl.
 26. The compound of the claim 25, whichis p-nitrobenzyl7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-cephem-4-carboxylate(syn isomer).
 27. An antibacterial composition comprising, as aneffective ingredient, the compound of claim 1, in association with apharmaceutically acceptable, substantially nontoxic carrier orexcipient.
 28. A method for treating an infectious disease caused bypathogens, which comprises administering the compound of claim 1 toinfected human being or animals.